Bone defects caused by severe trauma, tumors, infections and diseases remain a global challenge due to limited natural regeneration capacity of bone tissue in large-scale or complex injuries. Guided bone regeneration (GBR) has emerged as a pivotal technique in addressing these issues, relying on barrier membranes to facilitate osteoprogenitor cell infiltration. Current clinical GBR membranes function solely as physical barriers, lacking antibacterial and osteoinductive properties, which underscores the need for advanced alternatives. This study focuses on resorbable GBR membranes made from small intestinal submucosa (SIS), known for biocompatibility and tissue regeneration but hindered by low mechanical strength and rapid degradation. In addition, SIS lacks both antibacterial properties and strong osteogenic capabilities. Enhancements involve crosslinking treatment and dual incorporation of calcium (Ca2+) and zinc (Zn2+), which address the physical property shortcomings and synergistically boost osteoinductivity by activating osteogenic signaling pathways. Additionally, phase-transited lysozyme (PTL) nanofilm technique enables efficient ion loading and controlled release, while offering antibacterial properties. In this study, a multifunctional SIS membrane is constructed by PTL-ions layers, providing a potential solution to the challenge of clinical bone defects.

Tarsal coalition can be a long term severely disabling condition. For symptomatic cases with flatfoot surgical resection of coalition and subtalar arthroeresis represents the most common treatment. Literature reports variable outcomes and recurrence. The aim was to achieve optimal correction with no recurrence. This retrospective study presents, reporting results, an innovative technique that provides numerous advantages and improved outcomes. Nineteen patients suffering from painful flatfoot from tarsal coalition were, consecutively, surgically treated by resection of tarsal coalition with interposition of a synthetic flexible bioresorbable polymers membrane as an adhesion barrier and by subtalar arthroeresis. AOFAS scores were used to rate the clinical severity and X-ray/ CT to evaluate the extent of tarsal coalition. Patient's age at time of surgery ranged from 12 to 21. The period examined runs from November 2010 to November 2019. Results were evaluated (up to 9 years follow-up) clinically by AFOAS scores and radiologically by X-ray/CT. AOFAS scores improved in all patients with significant (p < 0.01) pain reduction or disappearance, corrected alignment and increased function, biomechanics of the foot and mobility. X-ray showed no recurrence of coalition in all but one case. There were no complications and patients reported a significant improvement in quality of life. Our study shows that surgical resection of coalition, and correction of flatfoot by subtalar arthroeresis, with the innovative use of a flexible bioresorbable polymers membrane as an adhesion barrier, obtained excellent overall results and importantly prevented recurrence. We believe this technique represents a great option. LEVEL OF CLINICAL EVIDENCE: 4.

Although allografts remain the gold standard for treating critical-size bone defects, ~60% fail within 10 years of implantation. To emulate periosteum-mediated healing of live autografts, we have developed a tissue-engineered periosteum (TEP) to improve allograft healing. The TEP comprises cell-degradable poly(ethylene glycol) hydrogels encapsulating mouse mesenchymal stem cells and osteoprogenitor cells to mimic the periosteal cell population. Despite improvements in allograft healing, several limitations were observed using the TEP, specifically the modulation of host tissue infiltration and remodeling to support graft-localized vascular volume and callus bridging. Therefore, hydrogel biochemical cues were incorporated into TEP to enable cell-matrix interactions and remodeling critical for tissue infiltration. Adhesive peptide functionalization (RGD, YIGSR, and GFOGER) and enzymatic degradation rate (GPQGIWGQ, IPESLRAG, and VPLSLYSG) were screened using an in vitro 3D cell spheroid assay and design of experiments (DOE) to identify hydrogels that best supported tissue infiltration and integration. DOE analysis of various adhesive peptide combinations was used to optimize functionalization, revealing that individual RGD-functionalization and GFOGER-functionalization maximized in vitro cell infiltration. RGD and GFOGER hydrogels were then investigated in vivo as TEP (RGD-TEP and GFOGER-TEP, respectively) to evaluate the effect of hydrogel functionalization on TEP-mediated allograft healing in a murine femur defect model. RGD- and GFOGER-TEP promoted bone graft healing, with both groups exhibiting a 1.9-fold increase in bone callus volume over unmodified allografts at 3 weeks post-implantation. RGD-TEP promoted more significant bone tissue development, but GFOGER-TEP promoted greater torsional biomechanics over time. The few differences observed between TEP groups suggest hydrogel functionalization has a limited effect on TEP-mediated healing, with cell delivery via the TEP enough to improve bone regeneration. Future studies aim to investigate additional adhesive peptides with diverse combinations to identify potential synergies between adhesive peptides to promote TEP-mediated bone allograft healing.

MXenes are a class of 2D transition metal carbides and nitrides (Mn+1XnT) that have attracted significant interest owing to their remarkable potential in various fields. The unique combination of their excellent electromagnetic, optical, mechanical, and physical properties have extended their applications to the biological realm as well. In particular, their ultra-thin layered structure holds specific promise for diverse biomedical applications. This comprehensive review explores the synthesis methods of MXene composites, alongside the biological and medical design strategies that have been employed for their surface engineering. This review delves into the interplay between these strategies and the resulting properties, biological activities, and unique effects at the nano-bio-interface. Furthermore, the latest advancements in MXene-based biomaterials and medicine are systematically summarized. Further discussion on MXene composites designed for various applications, including biosensors, antimicrobial agents, bioimaging, tissue engineering, and regenerative medicine, are also provided. Finally, with a focus on translating research results into real-world applications, this review addresses the current challenges and exciting future prospects of MXene composite-based biomaterials.

Periodontitis and bone loss in the maxillofacial and dental areas pose considerable challenges for both functional and aesthetic outcomes. To date, implantable dental barrier membranes, designed to prevent epithelial migration into defects and create a favorable environment for targeted cells, have garnered significant interest from researchers. Consequently, a variety of materials and fabrication methods have been explored in extensive research on regenerative dental barrier membranes.

This review focuses on dental barrier membranes, summarizing the various biomaterials used in membrane manufacturing, fabrication methods, and state-of-the-art applications for dental tissue regeneration. Based on a discussion of the pros and cons of current membrane strategies, future research directions for improved membrane designs are proposed.

To endow dental membranes with various biological properties that accommodate different clinical situations, numerous biomaterials and manufacturing methods have been proposed. These approaches provide theoretical support and hold promise for advancements in dental tissue regeneration.

Autograft has long been the gold standard for various bone surgeries. Nevertheless, the increasing usage of synthetic implants is taking over the operation rooms due to biosafety and standardized protocols. To fulfill such tremendous needs, a magnesium-impregnated membrane is devised that steadily releases magnesium ions to stimulate osteogenesis. The compatibility of Magnesium oxide (MgO) particles is enhanced through hydration and grafting, characterized by scanning electron microscopy (SEM), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). With detailed degradation profiles, an in-depth investigation of Magnesium transporter 1 (MagT1) for magnesium intake is carried out and engaging in the N-linked glycosylation by using RNAi and inhibitors. The glycosylation of secreted protein acidic and rich in cysteine (SPARC) affected extracellular secretion and mineral deposition, demonstrated by immunostaining and density-dependent color-SEM (DDC-SEM). Skull defects are treated by implanting magnesium-impregnated membranes in rats and evaluated them by micro-CT and histological exams. This study revealed the compatible integration of grafted magnesium hydroxide (g-MH) particles is the key to functional performance and critical to applicability in vivo; meanwhile, it opens the door to a biological rationale for designing biomimetic materials.

Collagen films play an essential role in guided bone-regeneration (GBR) techniques, which create space, promote cell adhesion, and induce osteogenic differentiation. It is therefore crucial to design appropriate GBR films to facilitate bone regeneration. However, current electrospun collagen scaffolds used as bioactive materials have limited clinical applications due to their poor mechanical properties. In this study, polyvinyl alcohol (PVA)/collagen (Col) films were electrospun by mixing PVA and type I collagen solution. For the first time, the intrafibrillar mineralization of aragonite nanocrystals within the PVA/Col fibrils was achieved, resulting in the formation of a hierarchical, bioactive film. The PVA/Col-CaCO3 film exhibited good mechanical properties, with hardness and Young's modulus values of 211.6 ± 0.1 MPa and 5.6 ± 1.7 GPa, respectively. Furthermore, bone marrow mesenchymal stem cells (BMSCs) inoculated onto the PVA/Col-CaCO3 film demonstrated robust adhesion and proliferation. The mineralized fibrils effectively stimulated the growth of BMSCs while suppressing cell apoptosis. Besides, the PVA/Col-CaCO3 film significantly induced the osteogenic differentiation of BMSCs, revealing its potential biomedical applications in hard tissue engineering.

(1) Background: Collagen, a natural polymer, is widely used in the fabrication of membranes for guided bone regeneration (GBR). These membranes are sourced from various tissues, such as skin, pericardium, peritoneum, and tendons, which exhibit differences in regenerative outcomes. Therefore, this study aimed to evaluate the morphological and chemical properties of porcine collagen membranes from five different tissue sources: skin, pericardium, dermis, tendons, and peritoneum. (2) Methods: The membrane structure was analyzed using energy-dispersive X-ray spectrometry (EDX), variable pressure scanning electron microscopy (VP-SEM), Fourier transform infrared spectroscopy (FTIR), and thermal stability via thermogravimetric analysis (TGA). The absorption capacity of the membranes for GBR was also assessed using an analytical digital balance. (3) Results: The membranes displayed distinct microstructural features. Skin- and tendon-derived membranes had rough surfaces with nanopores (1.44 ± 1.24 µm and 0.46 ± 0.1 µm, respectively), while pericardium- and dermis-derived membranes exhibited rough surfaces with macropores (78.90 ± 75.89 µm and 64.89 ± 13.15 µm, respectively). The peritoneum-derived membrane featured a rough surface of compact longitudinal fibers with irregular macropores (9.02 ± 3.70 µm). The thickness varied significantly among the membranes, showing differences in absorption capacity. The pericardium membrane exhibited the highest absorption, increasing by more than 10 times its initial mass. In contrast, the skin-derived membrane demonstrated the lowest absorption, increasing by less than 4 times its initial mass. Chemical analysis revealed that all membranes were primarily composed of carbon, nitrogen, and oxygen. Thermogravimetric and differential scanning calorimetry analyses showed no significant compositional differences among the membranes. FTIR spectra confirmed the presence of collagen, with characteristic peaks corresponding to Amide A, B, I, II, and III. (4) Conclusions: The tissue origin of collagen membranes significantly influences their morphological characteristics, which may, in turn, affect their osteogenic properties. These findings provide valuable insights into the selection of collagen membranes for GBR applications.

Guided bone regeneration (GBR) is an extensively used technique for the treatment of maxillofacial bone defects and bone mass deficiency in clinical practice. However, to date, studies on membranes for GBR have not achieved the combination of suitable properties and cost-effective membrane production. Herein, we developed a polycaprolactone/human extracellular matrix-like collagen (PCL/hCol) membrane with an asymmetric porous structure via the nonsolvent-induced phase separation (NIPS) method, which is a highly efficient procedure with simple operation, scalable fabrication and low cost. This membrane possessed a porous rough surface, which is conducive to cell attachment and proliferation for guiding osteogenesis, together with a relatively smooth surface with micropores, which allows the passage of nutrients and is unfavorable for the adhesion of cells, thus preventing fibroblast invasion and overall meeting the demands for GBR. Besides, we evaluated the characteristics and biological properties of the membrane and compared them with those of commercially available membranes. Results showed that the PCL/hCol membrane exhibited excellent mechanical properties, degradation characteristics, barrier function, biocompatibility and osteoinductive potential. Furthermore, our in vivo study demonstrated the promotive effect of the PCL/hCol membrane on bone formation in rat calvarial defects. Taken together, our NIPS-prepared PCL/hCol membrane with promising properties and production advantages offers a new perspective for its development and potential use in GBR application.

Wide used guided bone regeneration (GBR) membrane materials, such as collagen, Teflon, and other synthesized polymers, present a great challenge in term of integrating the mechanical property and degradation rate when addressing critical bone defects. Therefore, inspired by the distinctive architecture of fish scales, this study utilized epigallocatechin gallate to modify decellularized fish scales following biomimetic mineralization to fabricate a GBR membrane that mimics the structure of lamellar bone. The structure, physical and chemical properties, and biological functions of the novel GBR membrane were evaluated. Results indicate that the decellularized fish scale with 5 remineralization cycles (5R-E-DCFS) exhibited a composite and structure similar to natural bone and had a special functionally gradient mineral contents character, demonstrating excellent mechanical properties, hydrophilicity, and degradation properties. In vitro, the 5R-E-DCFS membrane exhibited excellent cytocompatibility promoting Sprague-Dawley (SD) rat bone marrow mesenchymal stem cell proliferation and differentiation up-regulating the expression of osteogenic-related genes and proteins. Furthermore, in vivo, the 5R-E-DCFS membrane promoted the critical skull bone defects of SD rats repairment and regeneration. Therefore, this innovative biomimetic membrane holds substantial clinical potential as an ideal GBR membrane with mechanical properties for space-making and suitable degradation rate for bone regeneration to manage bone defects.

Collagen has long been recognized as an excellent carrier for growth factors, and membrane-type collagen has been widely applied in dentistry for guided bone regeneration. This study was conducted to examine the effects of an activin A/BMP2 chimera (AB204) combined with a collagen membrane (CM) on bone repair in a rat calvarial defect model.

A unilateral calvarial defect measuring 5.0 mm was surgically created in 32 Sprague-Dawley rats. The rats were then randomly assigned to 1 of 4 groups, each consisting of 8 animals: control (untreated), CM (treated with a CM only), CM/bone morphogenetic protein 2 (BMP2) (treated with a CM and 1.0 μg of BMP2), and CM/AB204 (treated with a CM and 1.0 μg of AB204). Bone regeneration was evaluated using micro-computed tomography (CT) and histological analysis at 2 and 4 weeks following surgery.

Micro-CT analysis revealed that bone formation in the CM/BMP2 and CM/AB204 groups was superior to that observed in the control and CM groups at both 2 and 4 weeks postoperatively. BMP2 induced greater bone regeneration than AB204 at 2 weeks; however, AB204 resulted in a greater bone volume at 4 weeks, achieving the highest values recorded. No significant differences were found between the CM/BMP2 and CM/AB204 groups at either time point (P>0.05). On histological examination, new bone formation was evident in both CM/BMP2 and CM/AB204 groups.

Within the limitations of this study, the findings indicate that AB204 may enhance osteogenic potential when used in combination with CM for bone regeneration.

The present study aimed to investigate the histomorphometric and immunohistochemical impacts of vitamin K2 on guided bone regeneration (GBR) in calvarial critical-size defects (CSDs) in diabetic rats.

A total of 30 rats were used in this study, comprising 12 non-diabetic (control) rats and 18 with streptozotocin-nicotinamide-induced experimental Diabetes mellitus (DM). In all rats, two calvarial CSDs were created: one defect was left empty (E), the other was treated with bovine-derived bone graft and collagen-based resorbable membrane (GM). Study groups were as follows: control rats administered saline (n = 6, C-E and C-GM groups) or vitamin K2 (n = 6, CK-E and CK-GM groups) and diabetic rats administered saline (n = 6, DM-E and DM-GM groups) or vitamin K2 (n = 6, DMK-E and DMK-GM groups). After 4 weeks of saline or vitamin K2 administration, the rats were euthanized. Bone defect healing and new bone formation were assessed histomorphometrically, and osteocalcin and osteopontin levels were examined immunohistochemically.

Percentage of new bone formation was greater in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 3.86 (95% CI = 16.38-28.61), d = 1.86, (95% CI = 10.74-38.58), respectively, p < .05]. Bone defect healing scores were higher in CK-GM vs. CK-E and in DMK-GM vs. DMK-E [d = 2.69 (95% CI = -2.12 to -0.87), d = 3.28 (95% CI = 0.98-1.91), respectively, p < .05]. Osteocalcin expression levels were elevated in CK-GM vs. CK-E, in DMK-GM vs. DMK-E [d = 1.19 (95% CI = 0.08-1.41), d = 1.10 (95% CI = 0.02-1.22), respectively p < .05]. Vitamin K2 enhanced osteocalcin expression levels in DMK-E vs. DM-E [d = 2.78, (95% CI = 0.56-1.53), p < .05] and in DMK-GM vs. DM-GM [d = 2.43, (95% CI = 0.65-2.10), p < .05]. Osteopontin expression was enhanced in defects treated with GM vs. E defects [C-GM vs. C-E, d = 1.56 (95% CI = 0.38-2.01); CK-GM vs. CK-E, d = 1.91 (95% CI = 0.49-1.72); DM-GM vs. DM-E, d = 2.34 (95% CI = -1.12 to -0.50); DMK-GM vs. DMK-E, d = 2.00 (95% CI = 0.58-1.91), p < .05].

The research findings suggest that administering vitamin K2 in GBR for rats with DM favorably impacts bone healing in CSDs, presenting an adjunctive strategy for bone regeneration.

Periodontitis is a highly prevalent disease that damages the supporting tissues of a tooth, including the alveolar bone. Alveolar bone loss owing to periodontitis is broadly categorized as supra-alveolar and intra-alveolar bone loss. In intra-alveolar bone loss, the defect has an angular or oblique orientation to the long axis of the tooth in an apical direction. In contrast, the defect is perpendicular to the long axis of the tooth in supra-alveolar bone loss. Unlike intra-alveolar bone defects, supra-alveolar bone defects lack supporting adjacent space, which makes supra-alveolar bone regeneration more challenging. In addition, the limited availability of resources in terms of vascularity and underlying tissues is another obstacle to supra-alveolar bone regeneration. Currently, supra-alveolar bone loss is the least predictable periodontal defect type in regenerative periodontal therapy. In addition, supra-alveolar bone loss is much more common than other alveolar bone loss. Despite its prevalence, research on supra-alveolar bone regeneration remains sparse, indicating an unmet need for significant research efforts in this area. This review summarize recent advances, obstacles, and future directions in the field of supra-alveolar bone regeneration. We discuss the biomaterials, bioactive molecules, and cells that have been tested for supra-alveolar bone regeneration, followed by pre-clinical and clinical approaches employed in this field. Additionally, we highlight obstacles and present future directions that will propel supra-alveolar bone research forward.

Objectives: The technique of guided bone regeneration (GBR) has been widely used in the field of reconstructive dentistry to address hard tissue deficiency. The objective of this research was to manufacture a novel bi-layered asymmetric membrane that incorporates demineralized dentin matrix (DDM), a bioactive bone replacement derived from dentin, in order to achieve both soft tissue isolation and hard tissue regeneration simultaneously. Methods: DDM particles were harvested from healthy, caries-free permanent teeth. The electrospinning technique was utilized to synthesize bi-layered DDM-loaded PLGA/PLA (DPP) membranes. We analyzed the DPP bilayer membranes' surface topography, physicochemical properties and degradation ability. Rat skull critical size defects (CSDs) were constructed to investigate in vivo bone regeneration. Results: The synthesized DPP bilayer membranes possessed suitable surface characteristics, acceptable mechanical properties, good hydrophilicity, favorable apatite forming ability and suitable degradability. Micro-computed tomography (CT) showed significantly more new bone formation in the rat skull defects implanted with the DPP bilayer membranes. Histological evaluation further revealed that the bone was more mature with denser bone trabeculae. In addition, the DPP bilayer membrane significantly promoted the expression of the OCN matrix protein in vivo. Conclusions: The DPP bilayer membranes exhibited remarkable biological safety and osteogenic activity in vivo and showed potential as a prospective candidate for GBR applications in the future.

In the field of conservative dentistry and endodontics, mineral trioxide aggregate (MTA), commonly used, possesses advantages such as biocompatibility, antimicrobial properties and osteogenic potential. This study investigated the feasibility of utilizing membrane form mineral trioxide aggregate (MTA) as a barrier membrane in guided bone regeneration (GBR) procedures.

Membranes were electrospun from three different formulations: 15 w/v% Polycaprolactone (PCL), 13 w/v% PCL + 2 w/v% MTA (2MTA), and 11 w/v% PCL + 4 w/v% MTA (4MTA). Physicochemical and mechanical properties of the electrospun membrane were compared, encompassing parameters such as surface morphology, fiber diameter distribution, chemical composition, phase identification, tensile stress, pH variation, and water contact angle. Moreover, the antimicrobial properties against of the electrospun membranes were assessed through direct exposure to streptococcus aureus (S. aureus) and candida albicans (C. albicans). Additionally, on the 7th day, biocompatibility and cell attachment were investigated with respect to L929 (fibroblast) and MC3T3 (pre-osteoblast) cells. Inhibition of L929 cell infiltration and the expression of osteogenic related genes including osteocalcin (OCN), alkaline phosphatase (ALP), and runt related transcription factor 2 (RUNX2) in MC3T3 cells on 7th and 14th days were also investigated.

PCL, 2MTA, and 4MTA exhibited no statistically differences in fiber diameter distribution and tensile stress. However, as the MTA content increased, wettability and pH also increased. Due to the elevated pH, 4MTA demonstrated the lowest viability S.aureus and C.albicans. All membranes were highly biocompatibility and promoted cell attachment, while effectively preventing L929 cell infiltration. Lastly 4MTA showed increase in OCN, ALP, and RUNX2 expression on both 7th and 14th day.

The membrane form MTA possessed characteristics essential for a novel barrier membrane.

Guided bone regeneration (GBR) stands as an essential modality for craniomaxillofacial bone defect repair, yet challenges like mechanical weakness, inappropriate degradability, limited bioactivity, and intricate manufacturing of GBR membranes hindered the clinical efficacy. Herein, we developed a Janus bacterial cellulose(BC)/MXene membrane through a facile vacuum filtration and etching strategy. This Janus membrane displayed an asymmetric bilayer structure with interfacial compatibility, where the dense layer impeded cell invasion and the porous layer maintained stable space for osteogenesis. Incorporating BC with Ti3C2Tx MXene significantly enhanced the mechanical robustness and flexibility of the material, enabling clinical operability and lasting GBR membrane supports. It also contributed to a suitable biodegradation rate, which aligned with the long-term bone repair period. After demonstrating the desirable biocompatibility, barrier role, and osteogenic capability in vitro, the membrane's regenerative potential was also confirmed in a rat cranial defect model. The excellent bone repair performance could be attributed to the osteogenic capability of MXene nanosheets, the morphological cues of the porous layer, as well as the long-lasting, stable regeneration space provided by the GBR membrane. Thus, our work presented a facile, robust, long-lasting, and biodegradable BC/MXene GBR membrane, offering a practical solution to craniomaxillofacial bone defect repair.

Periodontitis, a persistent inflammatory condition, results in the deterioration of both the hard and soft tissues in the periodontium, leading to the formation of intrabony defects. Restoring the lost tissues, particularly bone, is possible through tissue engineering techniques utilizing scaffolds made from different polymers. Consequently, this research focuses on creating and assessing a scaffold infused with alginate (Sigma Aldrich, Gillingham, UK) and carrageenan (Sigma Aldrich, Gillingham, UK) for the purpose of bone regeneration.

An in vitro investigation was conducted to assess the characteristics of the recently formulated scaffold. Spectroscopic analysis, tensile strength testing, scanning electron microscopy (SEM) analysis, and degradation testing were carried out to evaluate both the physical and biological attributes of the scaffold.

IBM SPSS Statistics for Windows, V. 1.2 (IBM Corp., Armonk, NY, USA) was used for statistical analysis. A one-way ANOVA test was done to determine the significance of tensile strength, and a paired t-test was done to check the significance of the degradation test. The in vitro research unveiled notable distinctions in the physical and biological attributes between the scaffold infused with alginate and carrageenan and the PerioCol® (p<0.05).

The scaffold incorporating alginate and carrageenan demonstrated superior outcomes concerning parameters such as tensile stress and strain, degradation rate, percentage bone volume, and object surface density when contrasted with the conventional PerioCol®. Therefore, the scaffold infused with alginate and carrageenan emerges as a promising candidate for bone regeneration.

The effective repair of large bone defects remains a major challenge due to its limited self-healing capacity. Inspired by the structure and function of the natural periosteum, an electrospun biomimetic periosteum is constructed to programmatically promote bone regeneration using natural bone healing mechanisms. The biomimetic periosteum is composed of a bilayer with an asymmetric structure in which an aligned electrospun poly(ε-caprolactone)/gelatin/deferoxamine (PCL/GEL/DFO) layer mimics the outer fibrous layer of the periosteum, while a random coaxial electrospun PCL/GEL/aspirin (ASP) shell and PCL/silicon nanoparticles (SiNPs) core layer mimics the inner cambial layer. The bilayer controls the release of ASP, DFO, and SiNPs to precisely regulate the inflammatory, angiogenic, and osteogenic phases of bone repair. The random coaxial inner layer can effectively antioxidize, promoting cell recruitment, proliferation, differentiation, and mineralization, while the aligned outer layer can promote angiogenesis and prevent fibroblast infiltration. In particular, different stages of bone repair are modulated in a rat skull defect model to achieve faster and better bone regeneration. The proposed biomimetic periosteum is expected to be a promising candidate for bone defect healing.

Acquired cranial defects are a prevalent condition in neurosurgery and call for cranioplasty, where the missing or defective cranium is replaced by an implant. Nevertheless, the biomaterials in current clinical applications are hardly exempt from long-term safety and comfort concerns. An appealing solution is regenerative cranioplasty, where biomaterials with/without cells and bioactive molecules are applied to induce the regeneration of the cranium and ultimately repair the cranial defects. This review examines the current state of research, development, and translational application of regenerative cranioplasty biomaterials and discusses the efforts required in future research. The first section briefly introduced the regenerative capacity of the cranium, including the spontaneous bone regeneration bioactivities and the presence of pluripotent skeletal stem cells in the cranial suture. Then, three major types of biomaterials for regenerative cranioplasty, namely the calcium phosphate/titanium (CaP/Ti) composites, mineralised collagen, and 3D-printed polycaprolactone (PCL) composites, are reviewed for their composition, material properties, and findings from clinical trials. The third part discusses perspectives on future research and development of regenerative cranioplasty biomaterials, with a considerable portion based on issues identified in clinical trials. This review aims to facilitate the development of biomaterials that ultimately contribute to a safer and more effective healing of cranial defects.

Persistent bleeding and the absence of alveolar bone stress following tooth loss can hinder socket healing, complicating future dental implant procedures, and potentially leading to neighboring tooth instability. Therefore, developing materials that promote alveolar bone regeneration and possess both hemostatic and osteogenic properties is crucial for preserving the extraction sites. This study introduces a silk-based laponite composite scaffold material with proficient hemostatic and osteogenic functions, and excellent shape-memory properties for efficient extraction- site filling. In vitro studies research demonstrated that the scaffold's inherent negative charge of the scaffold significantly enhanced blood coagulation and thrombin generation. Moreover, its porous structure and slightly rough inner surface promoted blood cell adhesion and, improved the hemostatic performance. Furthermore, the scaffold facilitated stem cell osteogenic differentiation by activating the TRPM7 channel through the released of magnesium ions. In vivo tests using rat models confirmed its effectiveness in promoting coagulation and mandibular regeneration. Thus, this study proposes a promising approach for post-extraction alveolar bone regenerative repair. The composite scaffold material, with its hemostatic and osteogenic capabilities and shape-memory features, can potentially enhance dental implant success and overall oral health.

The implementation of a successful therapeutic approach that includes tissue-engineered grafts requires detailed analyses of graft-immune cell interactions in order to predict possible immune reactions after implantation. The phenotypic plasticity of macrophages plays a central role in immune cell chemotaxis, inflammatory regulation and bone regeneration. The present study addresses effects emanating from JPC-seeded β-TCP constructs (3DJPCs) co-cultivated with THP-1 derived M1/M2 macrophages within a horizontal co-culture system. After five days of co-culture, macrophage phenotype and chemokine secretion were analyzed by flow cytometry, quantitative PCR and proteome arrays. The results showed that pro-inflammatory factors in M1 macrophages were inhibited by 3DJPCs, while anti-inflammatory factors were activated, possibly affected by the multiple chemokines secreted by 3D-cultured JPCs. In addition, osteoclast markers of polarized macrophages were inhibited by osteogenically induced 3DJPCs. Functional assays revealed a significantly lower percentage of proliferating CD4+ T cells in the groups treated with secretomes from M1/M2 macrophages previously co-cultured with 3DJPCs compared to controls without secretomes. Quantifications of pit area resorption assays showed evidence that supernatants from 3DJPCs co-cultured with M1/M2 macrophages were able to completely suppress osteoclast maturation, compared to the control group without secretomes. These findings demonstrate the ability of 3D cultured JPCs to modulate macrophage plasticity.

Both guided bone and guided tissue regeneration are techniques that require the use of barrier membranes. Contamination and infection of the surgical area is one of the most feared complications. Some current lines of research focus on functionalizing these membranes with different antimicrobial agents. The objective of this study was to carry out a review of the use and antibacterial properties of regeneration membranes doped with antimicrobials such as zinc, silver, chlorhexidine, and lauric acid. The protocol was based on PRISMA recommendations, addressing the PICO question: "Do membranes doped with non-antibiotic antimicrobials have antibacterial activity that can reduce or improve infection compared to membranes not impregnated with said antimicrobial?" Methodological quality was evaluated using the RoBDEMAT tool. A total of 329 articles were found, of which 25 met the eligibility criteria and were included in this review. Most studies agree that zinc inhibits bacterial growth as it decreases colony-forming units, depending on the concentration used and the bacterial species studied. Silver compounds also decreased the secretion of proinflammatory cytokines and presented less bacterial adhesion to the membrane. Some concentrations of chlorhexidine that possess antimicrobial activity have shown high toxicity. Finally, lauric acid shows inhibition of bacterial growth measured by the disk diffusion test, the inhibition zone being larger with higher concentrations. Antimicrobial agents such as zinc, silver, chlorhexidine, and lauric acid have effective antibacterial activity and can be used to dope regenerative membranes in order to reduce the risk of bacterial colonization.

The utilization of guided tissue regeneration membranes is a significant approach for enhancing bone tissue growth in areas with bone defects. Biodegradable magnesium alloys are increasingly being used as guided tissue regeneration membranes due to their outstanding osteogenic properties. However, the degradation rates of magnesium alloy bone implants documented in the literature tend to be rapid. Moreover, many studies focus only on the initial 3-month period post-implantation, limiting their applicability and impeding clinical adoption. Furthermore, scant attention has been given to the interplay between the degradation of magnesium alloy implants and the adjacent tissues. To address these gaps, this study employs a well-studied magnesium-aluminum (Mg-Al) alloy membrane with a slow degradation rate. This membrane is implanted into rat skull bone defects and monitored over an extended period of up to 48 weeks. Observations are conducted at various intervals (2, 4, 8, 12, 24, and 48 weeks) following the implantation. Assessment of degradation behavior and tissue regeneration response is carried out using histological sections, micro-CT scans, and scanning electron microscopy (SEM). The findings reveal that the magnesium alloy membranes demonstrate remarkable biocompatibility and osteogenic capability over the entire observation duration. Specifically, the Mg-Al alloy membranes sustain their structural integrity for 8 weeks. Notably, their osteogenic ability is further enhanced as a corrosion product layer forms during the later stages of implantation. Additionally, our in vitro experiments employing extracts from the magnesium alloy display a significant osteogenic effect, accompanied by a notable increase in the expression of osteogenic-related genes. Collectively, these results strongly indicate the substantial potential of Mg-Al alloy membranes in the context of guided tissue regeneration.

Marine polysaccharides (MPs) are exceptional bioactive materials that possess unique biochemical mechanisms and pharmacological stability, making them ideal for various tissue engineering applications. Certain MPs, including agarose, alginate, carrageenan, chitosan, and glucan have been successfully employed as biological scaffolds in animal studies. As carriers of signaling molecules, scaffolds can enhance the adhesion, growth, and differentiation of somatic cells, thereby significantly improving the tissue regeneration process. However, the biological benefits of pure MPs composite scaffold are limited. Therefore, physical, chemical, enzyme modification and other methods are employed to expand its efficacy. Chemically, the structural properties of MPs scaffolds can be altered through modifications to functional groups or molecular weight reduction, thereby enhancing their biological activities. Physically, MPs hydrogels and sponges emulate the natural extracellular matrix, creating a more conducive environment for tissue repair. The porosity and high permeability of MPs membranes and nanomaterials expedite wound healing. This review explores the distinctive properties and applications of select MPs in tissue regeneration, highlighting their structural versatility and biological applicability. Additionally, we provide a brief overview of common modification strategies employed for MP scaffolds. In conclusion, MPs have significant potential and are expected to be a novel regenerative material for tissue engineering.

Silk fibroin hydrogel is a widely used material for tissue engineering. However, its mechanical properties are the main obstacle to its application to medical bone regeneration barrier membranes. Here, we developed a new hydrogel membrane for guided bone regeneration (GBR). In this study, a sequential crosslinking strategy including photo crosslinking and organic solvent (ethanol) treatment was used to obtain silk fibroin hydrogel membrane (EA-SF). The mechanical properties of EA-SF were significantly enhanced compared to the hydrogel prepared only by photocrosslinking (E-SF). The compressive and tensile strengths of the hydrogel film treated with 75% ethanol for 6 h were 1.18 ± 0.36 MPa and 0.43 ± 0.03 MPa, respectively. In vitro cell culture results showed that EA-SF has good biocompatibility and can effectively shield fibroblasts (L929). EA-SF also has excellent in vitro protein hydrolysis stability. In vivo experiments (subcutaneous implantation and calvarial defect experiment) confirmed the stability and barrier functionality of EA-SF. In conclusion, this study demonstrated that ethanol could improve the mechanical properties of silk fibroin hydrogel and extend the scope of their application, making silk fibroin hydrogel a promising GBR material.

To evaluate the cellular response of both an intact fish skin membrane and a porcine-derived collagen membrane and investigate the bone healing response of these membranes using a translational, preclinical, guided-bone regeneration (GBR) canine model. Two different naturally sourced membranes were evaluated in this study: (i) an intact fish skin membrane (Kerecis Oral®, Kerecis) and (ii) a porcine derived collagen (Mucograft®, Geistlich) membrane, positive control. For the in vitro experiments, human osteoprogenitor (hOP) cells were used to assess the cellular viability and proliferation at 24, 48, 72, and 168 h. ALPL, COL1A1, BMP2, and RUNX2 expression levels were analyzed by real-time PCR at 7 and 14 days. The preclinical component was designed to mimic a GBR model in canines (n = 12). The first step was the extraction of premolars (P1-P4) and the 1st molars bilaterally, thereby creating four three-wall box type defects per mandible (two per side). Each defect site was filled with bone grafting material, which was then covered with one of the two membranes (Kerecis Oral® or Mucograft®). The groups were nested within the mandibles of each subject and membranes randomly allocated among the defects to minimize potential site bias. Samples were harvested at 30-, 60-, and 90-days and subjected to computerized microtomography (μCT) for three-dimensional reconstruction to quantify bone formation and graft degradation, in addition to histological processing to qualitatively analyze bone regeneration. Neither the intact fish skin membrane nor porcine-based collagen membrane presented cytotoxic effects. An increase in cell proliferation rate was observed for both membranes, with the Kerecis Oral® outperforming the Mucograft® at the 48- and 168-hour time points. Kerecis Oral® yielded higher ALPL expression relative to Mucograft® at both 7- and 14-day points. Additionally, higher COL1A1 expression was observed for the Kerecis Oral® membrane after 7 days but no differences were detected at 14 days. The membranes yielded similar BMP2 and RUNX2 expression at 7 and 14 days. Volumetric reconstructions and histologic micrographs indicated gradual bone ingrowth along with the presence of particulate bone grafts bridging the defect walls for both Kerecis Oral® and Mucograft® membranes, which allowed for the reestablishment of the mandible shape after 90 days. New bone formation significantly increased from 30 to 60 days, and from 60 to 90 days in vivo, without significant differences between membranes. The amount of bovine grafting material (%) within the defects significantly decreased from 30 to 90 days. Collagen membranes led to an upregulation of cellular proliferation and adhesion along with increased expression of genes associated with bone healing, particularly the intact fish skin membrane. Despite an increase in the bone formation rate in the defect over time, there was no significant difference between the membranes.

Non-resorbable dental barrier membranes entail the risk of dehiscence due to their smooth and functionally inert surfaces. Non-thermal plasma (NTP) treatment has been shown to increase the hydrophilicity of a biomaterials and could thereby enhance cellular adhesion. This study aimed to elucidate the role of allyl alcohol NTP treatment of poly(tetrafluoroethylene) in its cellular adhesion. The materials (non-treated PTFE membranes (NTMem) and NTP-treated PTFE membranes (PTMem)) were subjected to characterization using scanning electron microscopy (SEM), contact angle measurements, X-ray photoelectron spectroscopy (XPS), and electron spectroscopy for chemical analysis (ESCA). Cells were seeded upon the different membranes, and cellular adhesion was analyzed qualitatively and quantitatively using fluorescence labeling and a hemocytometer, respectively. PTMem exhibited higher surface energies and the incorporation of reactive functional groups. NTP altered the surface topography and chemistry of PTFE membranes, as seen through SEM, XPS and ESCA, with partial defluorination and polymer chain breakage. Fluorescence labeling indicated significantly higher cell populations on PTMem relative to its untreated counterparts (NTMem). The results of this study support the potential applicability of allyl alcohol NTP treatment for polymeric biomaterials such as PTFE-to increase cellular adhesion for use as dental barrier membranes.

Guided bone regeneration (GBR) is a widely used technique in preclinical and clinical studies due to its predictability. Its main purpose is to prevent the migration of soft tissue into the osseous wound space, while allowing osseous cells to migrate to the site. GBR is classified into two main categories: resorbable and non-resorbable membranes. Resorbable membranes do not require a second surgery but tend to have a short resorption period. Conversely, non-resorbable membranes maintain their mechanical strength and prevent collapse. However, they require removal and are susceptible to membrane exposure. GBR is often used with bone substitute graft materials to fill the defect space and protect the bone graft. The membrane can also undergo various modifications, such as surface modification and biological factor loading, to improve barrier functions and bone regeneration. In addition, bone regeneration is largely related to osteoimmunology, a new field that focuses on the interactions between bone and the immune system. Understanding these interactions can help in developing new treatments for bone diseases and injuries. Overall, GBR has the potential to be a powerful tool in promoting bone regeneration. Further research in this area could lead to advancements in the field of bone healing. This review will highlight resorbable and non-resorbable membranes with cellular responses during bone regeneration, provide insights into immunological response during bone remodeling, and discuss antibacterial features.

Dental implants revolutionized the treatment options for restoring form, function, and esthetics when one or more teeth are missing. At sites of insufficient bone, guided bone regeneration (GBR) is performed either prior to or in conjunction with implant placement to achieve a three-dimensional prosthetic-driven implant position. To date, GBR is well documented, widely used, and constitutes a predictable and successful approach for lateral and vertical bone augmentation of atrophic ridges. Evidence suggests that the use of barrier membranes maintains the major biological principles of GBR. Since the material used to construct barrier membranes ultimately dictates its characteristics and its ability to maintain the biological principles of GBR, several materials have been used over time. This review, summarizes the evolution of barrier membranes, focusing on the characteristics, advantages, and disadvantages of available occlusive barrier membranes and presents results of updated meta-analyses focusing on the effects of these membranes on the overall outcome.

Guided tissue regeneration (GTR) is an important surgical method for periodontal regeneration. By placing barrier membrane on the root surface of the tooth to guide the adhesion and proliferation of periodontal ligament cells, periodontal tissue regeneration can be achieved. This review intends to analyze the current limitations of GTR membranes and to propose possible solutions for developing new ones. Limitations of current GTR membranes include nonabsorbable membranes and absorbable synthetic polymer membranes exhibit weak biocompatibility; when applying to a large defect wound, the natural collagen membrane with fast degradation rate have limited mechanical strength, and the barrier function may not be maintained well. Although the degradation time can be prolonged after cross-linking, it may cause foreign body reaction and affect tissue integration; The clinical operation of current barrier membranes is inconvenient. In addition, most of the barrier membranes lack bioactivity and will not actively promote periodontal tissue regeneration. Possible solutions include using electrospinning (ELS) techniques, nanofiber scaffolds, or developing functional gradient membranes to improve their biocompatibility; adding Mg, Zn, and/or other metal alloys, or using 3D printing technology to improve their mechanical strength; increasing the concentration of nanoparticles or using directional arrangement of membrane fibers to control the fiber diameter and porosity of the membrane, which can improve their barrier function; mixing natural and synthetic polymers as well as other biomaterials with different degradation rates in proportion to change the degradation rate and maintain barrier function; to improve the convenience of clinical operation, barrier membranes that meets personalized adhesion to the wound defect can be manufactured; developing local controlled release drug delivery systems to improve their bioactivity. Impact statement This review provides an up-to-date summary of commonly commercial periodontal guided tissue regeneration membranes, and analyze their limitations in clinical use. Using studies published recently to explore possible solutions from several perspectives and to raise possible strategies in the future. Several strategies have tested in vivo/in vitro, which will guide the way to propel clinical translation, meeting clinical needs.

Critical-sized bone defects resulting from trauma, inflammation, and tumor resections are individual in their size and shape. Implants for the treatment of such defects have to consider biomechanical and biomedical factors, as well as the individual conditions within the implantation site. In this context, 3D printing technologies offer new possibilities to design and produce patient-specific implants reflecting the outer shape and internal structure of the replaced bone tissue. The selection or modification of materials used in 3D printing enables the adaption of the implant, by enhancing the osteoinductive or biomechanical properties. In this study, scaffolds with bone spongiosa-inspired structure for extrusion-based 3D printing were generated. The computer aided design process resulted in an up scaled and simplified version of the bone spongiosa. To enhance the osteoinductive properties of the 3D printed construct, polycaprolactone (PCL) was combined with 20% (wt) calcium phosphate nano powder (CaP). The implants were designed in form of a ring structure and revealed an irregular and interconnected porous structure with a calculated porosity of 35.2% and a compression strength within the range of the natural cancellous bone. The implants were assessed in terms of biocompatibility and osteoinductivity using the osteosarcoma cell line MG63 and patient-derived mesenchymal stem cells in selected experiments. Cell growth and differentiation over 14 days were monitored using confocal laser scanning microscopy, scanning electron microscopy, deoxyribonucleic acid (DNA) quantification, gene expression analysis, and quantitative assessment of calcification. MG63 cells and human mesenchymal stem cells (hMSC) adhered to the printed implants and revealed a typical elongated morphology as indicated by microscopy. Using DNA quantification, no differences for PCL or PCL-CaP in the initial adhesion of MG63 cells were observed, while the PCL-based scaffolds favored cell proliferation in the early phases of culture up to 7 days. In contrast, on PCL-CaP, cell proliferation for MG63 cells was not evident, while data from PCR and the levels of calcification, or alkaline phosphatase activity, indicated osteogenic differentiation within the PCL-CaP constructs over time. For hMSC, the highest levels in the total calcium content were observed for the PCL-CaP constructs, thus underlining the osteoinductive properties.

This comprehensive review provides an in-depth analysis of the use of biomaterials in the processes of guided tissue and bone regeneration, and their indispensable role in dental therapeutic interventions. These interventions serve the critical function of restoring both structural integrity and functionality to the dentition that has been lost or damaged. The basis for this review is laid through the exploration of various relevant scientific databases such as Scopus, PubMed, Web of science and MEDLINE. From a meticulous selection, relevant literature was chosen. This review commences by examining the different types of membranes used in guided bone regeneration procedures and the spectrum of biomaterials employed in these operations. It then explores the manufacturing technologies for the scaffold, delving into their significant impact on tissue and bone regenerations. At the core of this review is the method of guided bone regeneration, which is a crucial technique for counteracting bone loss induced by tooth extraction or periodontal disease. The discussion advances by underscoring the latest innovations and strategies in the field of tissue regeneration. One key observation is the critical role that membranes play in guided reconstruction; they serve as a barrier, preventing the entry of non-ossifying cells, thereby promoting the successful growth and regeneration of bone and tissue. By reviewing the existing literature on biomaterials, membranes, and scaffold manufacturing technologies, this paper illustrates the vast potential for innovation and growth within the field of dental therapeutic interventions, particularly in guided tissue and bone regeneration.

To provide a reference for clinical selection of collagen membranes by analyzing the properties of three kinds of collagen membranes widely used in clinics: Bio-Gide membrane from porcine dermis (PD), Heal-All membrane from bovine dermis (BD), and Lyoplant membrane from bovine pericardium (BP).

The barrier function of three kinds of collagen membranes were evaluated by testing the surface morphology, mechanical properties, hydrophilicity, and degradation rate of collagen membranes in collagenase and artificial saliva. In addition, the bioactivity of each collagen membrane as well as the proliferation and osteogenesis of MC3T3-E1 cells were evaluated. Mass spectrometry was also used to analyze the degradation products.

The BP membrane had the highest tensile strength and Young's modulus as well as the largest water contact angle. The PD membrane exhibited the highest elongation at break, the smallest water contact angle, and the lowest degradation weight loss. The BD membrane had the highest degradation weight loss, the highest number of proteins in its degradation product, the strongest effect on the proliferation of MC3T3-E1 cells, and the highest expression level of osteogenic genes.

The PD membrane is the best choice for shaping and maintenance time, while the BD membrane is good for osteogenesis, and the BP membrane is suitable for spatial maintenance. To meet the clinical requirements of guided bone regeneration, using two different kinds of collagen membranes concurrently to exert their respective advantages is an option worth considering.

Natural polymers are increasingly being used in tissue engineering due to their ability to mimic the extracellular matrix and to act as a scaffold for cell growth, as well as their possible combination with other osteogenic factors, such as mesenchymal stem cells (MSCs) derived from dental pulp, in an attempt to enhance bone regeneration during the healing of a bone defect. Therefore, the aim of this study was to analyze the repair of mandibular defects filled with a new collagen/chitosan scaffold, seeded or not with MSCs derived from dental pulp. Twenty-eight rats were submitted to surgery for creation of a defect in the right mandibular ramus and divided into the following groups: G1 (control group; mandibular defect with clot); G2 (defect filled with dental pulp mesenchymal stem cells-DPSCs); G3 (defect filled with collagen/chitosan scaffold); and G4 (collagen/chitosan scaffold seeded with DPSCs). The analysis of the scaffold microstructure showed a homogenous material with an adequate percentage of porosity. Macroscopic and radiological examination of the defect area after 6 weeks post-surgery revealed the absence of complete repair, as well as absence of signs of infection, which could indicate rejection of the implants. Histomorphometric analysis of the mandibular defect area showed that bone formation occurred in a centripetal fashion, starting from the borders and progressing towards the center of the defect in all groups. Lower bone formation was observed in G1 when compared to the other groups and G2 exhibited greater osteoregenerative capacity, followed by G4 and G3. In conclusion, the scaffold used showed osteoconductivity, no foreign body reaction, malleability and ease of manipulation, but did not obtain promising results for association with DPSCs.

Guided bone regeneration (GBR) utilizing eggshell membrane (ESM) as a potential biomaterial for dental implant therapy augmentation was explored in this study. ESM, an environmentally friendly waste product, possesses collagen-rich characteristics. The biocompatibility and histological responses of ESM were investigated in a rat model. Twelve young adult Wistar rats were used in this study. ESM samples were implanted in subcutaneous and intramuscular pockets, and samples were collected at 48 hours, 4 weeks, and 8 weeks post-implantation. Histological analysis revealed the changes in ESM over time. Results showed that ESM maintained its structural integrity, induced a moderate cellular response, and exhibited slow degradation, indicating potential biocompatibility. However, the lack of organized collagen arrangement in ESM led to the formation of irregular and polymorphic spaces, allowing cell migration. Encapsulation of ESM by newly proliferating collagen fibers and multinucleated giant cells was observed at later time points, indicating a foreign body reaction. Crosslinking might improve its performance as a separation membrane, as it has the potential to resist enzymatic degradation and enhance biomechanical properties. In conclusion, ESM demonstrated biocompatibility, slow degradation, and lack of foreign body reaction. While not suitable as a complete separation membrane due to irregular collagen arrangement, further research involving crosslinking could enhance its properties, making it a viable option for guided bone regeneration applications in dental implant therapy. This study highlights the potential of repurposing waste materials for medical purposes and underscores the importance of controlled collagen structure in biomaterial development.

The objective of this study was to evaluate the efficacy of photobiomodulation in the bone regeneration of critical-sized defects (CSD) filled with inorganic bovine bone associated or not with collagen membranes. The study has been conducted on 40 critical defects in the calvaria of male rats, divided into four experimental groups (n = 10): (1) DBBM (deproteinized bovine bone mineral); (2) GBR (DBBM+collagen membrane); (3) DBBM+P (DBBM+photobiomodulation); and (4) GBR+P (GBR+photobiomodulation). At 30 days postoperative, the animals were euthanized, and after the tissue had been processed, histological, histometric, and statistical analyses were performed. The analyses have taken into account newly formed bone area (NBA), linear bone extension (LBE), and residual particle area (RPA) as variables. The Kruskal-Wallis test has been performed, followed by the Dwass-Steel-Critchlow-Fligner test for comparison between groups (p < 0.05). When the DBBM+P group was compared to the DBBM group, it was possible to observe significant statistical differences in all the variables analyzed (p < 0.05). The application of photobiomodulation in guided bone regeneration (GBR+P) has shown a decrease in the median value for the RPA variable (26.8) when compared to the GBR group (32.4), with a significant statistical difference; however, for NBA and LBE, the therapy has not provided significant results.

Bioresorbable polymeric membranes for guided bone regeneration (GBR) were fabricated using the three-dimensional printing technique. Membranes made of polylactic-co-glycolic acid (PLGA), which consist of lactic acid (LA) and glycolic acid in ratios of 10:90 (group A) and 70:30 (group B), were compared. Their physical characteristics including architecture, surface wettability, mechanical properties, and degradability were compared in vitro, and their biocompatibilities were compared in vitro and in vivo. The results demonstrated that the membranes of group B had mechanical strength and could support the proliferation of fibroblasts and osteoblasts significantly better than those of group A (p < 0.05). The degradation rate in Group B was significantly lower than that in Group A, but they significantly produced less acidic environment (p < 0.05). In vivo, the membranes of group B were compared with the commercially available collagen membranes (group C). The amount of newly formed bone of rat's calvarial defects covered with the membranes of group C was stable after week 2, whereas that of group B increased over time. At week 8, the new bone volumes in group B were greater than those in group C (p > 0.05). In conclusion, the physical and biological properties of the PLGA membrane (LA:GA, 70:30) were suitable for GBR.

The Masquelet technique is a relatively new method for large bone defect treatment. In this technique, grafted bone tissue is used, and after the cement is removed, the induced membrane (IM; that form around the cement spacers placed in the bone defect region) is thought to play an important role in promoting bone formation. On the other hand, low-intensity pulsed ultrasound (LIPUS) is known to promote fracture healing and angiogenesis through mechanical stimulation. This study aimed to investigate the in vitro effects of LIPUS on the osteogenic differentiation of human induced membrane-derived cells (IMCs).

Seven patients who had been treated using the Masquelet technique were enrolled. The IM was harvested during the second stage of the technique. IMCs were isolated, cultured in growth medium, and then divided into two groups: (1) control group, IMCs cultured in osteogenic medium without LIPUS, and (2) LIPUS group, IMCs cultured in osteogenic medium with LIPUS treatment. Adherent cells from the IM samples were harvested after the first passage and evaluated for cell surface protein expression using immunostaining. A cell proliferation assay was used to count the number of IMCs using a hemocytometer. Osteogenic differentiation capability was assessed using an alkaline phosphatase (ALP) activity assay, Alizarin Red S staining, and real-time reverse transcription-polymerase chain reaction.

Cell surface antigen profiling revealed that the IMCs contained cells positive for the mesenchymal stem cell-related markers CD73, CD90, and CD105. No significant difference in cell numbers was found between the control and LIPUS groups. The ALP activity of IMCs in the LIPUS group was significantly higher than that in the control group on days 7 and 14. Alizarin red S staining intensity was significantly higher in the LIPUS group than in the control group on day 21. Runx2 and VEGF expression was significantly upregulated on days 7 and 14, respectively, compared with levels in the control group.

We demonstrated the significant effect of LIPUS on the osteogenic differentiation of human IMCs. This study indicates that LIPUS can be used as an additional tool for the enhancement of the healing process of the Masquelet technique.

Different collagen barrier membranes come in various sources and crosslinking that may affect barrier function and tissue integration. This study investigated barrier function and tissue integration of the three different collagen membranes (Jason®: porcine pericardium, GENOSS: bovine tendon, and BioMend® Extend: cross-linked bovine tendon) with human gingival fibroblasts. The barrier function and tissue integration properties were determined under confocal microscopy. Morphological characteristics were observed using scanning electron microscopy. Our results showed that all collagen membranes allowed a small number of cells to migrate, and the difference in barrier function ability was not significant. The cross-linked characteristics did not improve barrier ability. The native collagen membrane surfaces allowed evenly scattered proliferation of HGF, while the cross-linked collagen membrane induced patchy proliferation. Statistically significant differences in cell proliferation were found between Jason and BioMend Extend membranes (p = 0.04). Scanning electron microscope showed a compact membrane surface at the top, while the bottom surfaces displayed interwoven collagen fibers, which were denser in the crosslinked collagen membranes. Within the limitations of this study, collagen membranes of different origins and physical properties can adequately prevent the invasion of unwanted cells. Native collagen membranes may provide a better surface for gingival cell attachment and proliferation.

The current study aimed to perform an in vivo examination using a critical-size periodontal canine model to investigate the capability of a 3D-printed soft membrane for guided tissue regeneration (GTR). This membrane is made of a specific composition of gelatin, elastin, and sodium hyaluronate that was fine-tuned and fully characterized in vitro in our previous study. The value of this composition is its potential to be employed as a suitable replacement for collagen, which is the main component of conventional GTR membranes, to overcome the cost issue with collagen.

Critical-size dehiscence defects were surgically created on the buccal surface of the roots of canine bilateral mandibular teeth. GTR treatment was performed with the 3D-printed membrane and two commercially available collagen membranes (Botiss Jason® and Smartbrane-Regedent membranes) and a group without any membrane placement was considered as the control group. The defects were submerged with tension-free closure of the gingival flaps. Histologic and histometric analyses were employed to assess the periodontal healing over an 8-week experimental period.

Histometric evaluations confirmed higher levels of new bone formation in the 3D-printed membrane group. Moreover, in all defects treated with the membranes, the formation of periodontal tissues, bone, periodontal ligaments, and cementum was observed after 8 weeks, while in the control group, only connective tissue was found in the defect sites. There was no clinical sign of inflammation or recession of gingiva in any of the groups.

The 3D-printed gelatin/elastin/sodium hyaluronate membrane can be safe and effective for use in GTR for periodontal tissue regeneration therapies, with better or comparable results to the commercial collagen membranes.

To obtain multifunctional materials suitable for guiding alveolar bone regeneration under infectious conditions, we prepared asymmetric membranes comprising space acquiring layer that involves fibroblast inhibitor poly(p-dioxanone-co-L-phenylalanine) (PDPA), an isolating dense layer that forms barrier between two layers and an osteogenesis inducing electrospinning layer which involves hydroxyapatite or hydroxyapatite & minocycline. Then the composition, crystallization, morphology, and hydrophilicity of asymmetric membranes were analyzed. Minocycline incorporated membranes controlled the expansion of Porphyromonas gingivalis (P. gingivalis) in vitro. Hydroxyapatite-incorporated asymmetric membranes promoted the expression of osteogenesis related genes RUNX2, OPN, ALP of MC3T3-E1 cells in vitro. The mineralization of MC3T3-E1 cells cultured with hydroxyapatite-incorporated asymmetric membranes were also promoted in vitro. Asymmetric membranes especially hydroxyapatite-incorporated ones guided the regeneration of the mandibular bone defect in vivo. Bone regeneration guided under infectious conditions was evaluated in a P. gingivalis infected alveolar bone defect model. Specifically, space acquiring layer containing asymmetric membranes effectively controlled connective tissue hyperplasia at defect sites. The excellent guided bone regeneration achieved by applying a single space acquiring layer membrane further indicates the importance of acquiring space actively to induce bone regeneration. Hydroxyapatite-minocycline incorporated symmetric membranes could simultaneously suppress alveolar bone reabsorption caused by infection and guide regeneration of defects. Therefore, the hydroxyapatite-minocycline incorporated asymmetric membrane may be more suitable to be applied in guiding regeneration of bone defects under complex infectious conditions.

Non-resorbable PTFE membranes are frequently used in dental-guided bone regeneration (GBR). However, there is a lack of detailed comparative studies that define variations among commonly used PTFE membranes in daily dental clinical practice. The aim of this study was to examine differences in physicochemical and mechanical properties of several recent commercial PTFE membranes for dental GBR (CytoplastTM TXT-200, permamem®, NeoGen®, Surgitime, OsseoGuard®-TXT, OsseoGuard®-NTXT). Such differences have been rarely recorded so far, which might be a reason for the varied clinical results. For that reason, we analyzed their surface architecture, chemical composition, tensile strength, Young's modulus, wettability, roughness, density, thickness and porosity. SEM revealed different microarchitectures among the non-textured membranes; the textured ones had hexagonal indentations and XPS indicated an identical spectral portfolio in all membranes. NeoGen® was determined to be the strongest and OsseoGuard®-TXT was the most elastic. Wettability and roughness were highest for Surgitime but lowest for OsseoGuard®-NTXT. Furthermore, permamem® was the thinnest and NeoGen® was identified as the thickest investigated GBR membrane. The defect volumes and defect volume ratio (%) varied significantly, indicating that permamem® had the least imperfect structure, followed by NeoGen® and then Cytoplast TM TXT-200. These differences may potentially affect the clinical outcomes of dental GBR procedures.