Antimicrobial drugs of several classes play an important role in the treatment of bone and joint infections. In addition to fighting pathogenic microorganisms, the effects of drugs on local tissues and cells are also related to the course and prognosis of bone and joint infections. The multi-directional differentiation potential of bone marrow-derived mesenchymal stem cells (MSCs) is essential for tissue repair after local injury, which is directly related to the recovery of bone, cartilage, and medullary adipose tissue. Our previous studies and the literature indicate that certain antimicrobial agents can regulate the differentiation potential of bone marrow-derived MSCs. Here, in order to systematically analyze the effects of various antimicrobial drugs on local tissue regeneration, we comprehensively review the studies on the effects of these drugs on MSC differentiation, and classify them according to the three differentiation directions (osteogenesis, chondrogenesis, and adipogenesis). Our review demonstrates the specific effects of different antimicrobial agents on bone marrow-derived MSCs and the range of concentrations at which they work, and provides a basis for drug selection at different sites of infection.

Strategies to improve cartilage repair tissue quality after bone marrow cell-based procedures may reduce later development of osteoarthritis. Doxycycline is inexpensive, well-tolerated, and has been shown to reduce matrix-metalloproteinases (MMPs) and osteoarthritis progression. This study tests the hypotheses that doxycycline reduces MMP, enhances chondrogenesis of human bone marrow-derived mesenchymal stem cells (hMSC), and improves in vivo cartilage repair.

Ninety hMSC pellets were cultured in chondrogenic media with either 0-, 1- or 2-μg/mL doxycycline. Pellets were evaluated with stereomicroscopy, proteoglycan assay, qRT-PCR, and histology. Osteochondral defects (OCDs) were created in the trochlear grooves of 24-Sprague-Dawley rats treated with/without oral doxycycline. Rats were sacrificed at 12-weeks and repair tissues were examined grossly and histologically.

hMSC pellets with 1-μg/mL (P = 0.014) and 2-μg/mL (P = 0.002) doxycycline had larger areas than pellets without doxycycline. hMSC pellets with 2-μg/mL doxycycline showed reduced mmp-13 mRNA (P = 0.010) and protein at 21-days. Proteoglycan, DNA contents, and mRNA expressions of chondrogenic genes were similar (P > 0.05). For the in vivo study, while the histological scores were similar between the two groups (P = 0.116), the gross scores of the OCD repair tissues in doxycycline-treated rats were higher at 12-weeks (P = 0.017), reflective of improved repair quality. The doxycycline-treated repairs also showed lower MMP-13 protein (P = 0.029).

This study shows that doxycycline improves hMSC chondrogenesis and decreases MMP-13 in pellet cultures and within rat OCDs. Doxycycline exerted no negative effect on multiple measures of chondrogenesis and cartilage repair. These data support potential use of doxycycline to improve cartilage repair to delay the onset of osteoarthritis.

Although results of bone marrow stimulation in osteochondral defects of the talus (OCLT) have been satisfactory, the technique performance has not yet been subjected to review as a prognostic factor. The aim of this systematic review is to determine whether variation within technique influences outcome of bone marrow stimulation for OCLT. Electronic databases were searched for articles on OCLT treated with bone marrow stimulation techniques, providing a technique description. Six articles on microfracture were included (198 patients). Lesion size averaged 0.9 cm (2) to 4.5 cm (2), and follow-up varied from 2 to 6 years. Key elements were removal of unstable cartilage, hole depth variation between 2 and 4 mm until bleeding or fat droplets occurred, and a distance between the created holes of 3 to 4 mm. The success rate (excellent/good results by any clinical outcome score) was 81%. There is a vast similarity in the technique with similar outcomes as in previous general reviews; therefore variation in technique as currently described in the literature does not seem to influence the outcome of bone marrow stimulation for OCLT. Whether the instruments used or the hole depth and geometry influence clinical outcome remains to be determined. Microfracture is safe and effective for OCLTs smaller than 15 mm. However, in this review, only 81% of patients obtained satisfactory results. Larger clinical trials are needed with clearly defined patient groups, technique descriptions, and reproducible outcome measures to provide insight in the specific indications and the preferred technique of bone marrow stimulation.

OBJECTIVE: To test the hypothesis that platelet-rich fibrin glue (PR-FG) can be used clinically as a scaffold to deliver autologous culture-expanded bone marrow mesenchymal stem cells (BM-MSCs) for cartilage repair and to report clinical results 1 y after implantation of MSCs PR-FG. PATIENTS AND METHODS: Autologous BM-MSCs were culture expanded, placed on PR-FG intraoperatively, and then transplanted into 5 full-thickness cartilage defects of femoral condyles of 5 patients and covered with an autologous periosteal flap. Patients were evaluated clinically at 6 and 12 mo by the Lysholm and Revised Hospital for Special Surgery Knee (RHSSK) scores and radiographically by x-rays and magnetic resonance imaging (MRI) at the same time points. Repair tissue in 2 patients was rated arthroscopically after 12 mo using the International Cartilage Repair Society (ICRS) Arthroscopic Score. STUDY DESIGN: Case series; level of evidence 4. RESULTS: All patients' symptoms improved over the follow-up period of 12 mo. Average Lysholm and RHSSK scores for all patients showed statistically significant improvement at 6 and 12 mo postoperatively (P < 0.05). There was no statistically significant difference between the 6 and 12 mo postoperative clinical scores (P = 0.18). ICRS arthroscopic scores were 8/12 and 11/12 (nearly normal) for the 2 patients who consented to arthroscopy. MRI of 3 patients at 12 mo postoperatively revealed complete defect fill and complete surface congruity with native cartilage, whereas that of 2 patients showed incomplete congruity. CONCLUSION: Autologous BM-MSC transplantation on PR-FG as a cell scaffold may be an effective approach to promote the repair of articular cartilage defects of the knee in human patients.

Chondrocytes depend on their environment to aid in their expression of appropriate proteins. It has been found that the interaction of integrin receptors with chondrocytes effects the production of extracellular molecules such as type II collagen and aggrecan. Additionally, the presence of growth factors such as IGF-1, TGF-beta1 and BMP-7 induce various signaling pathways that also aid in transducing phenotypic expressions by chondrocytes. Natural and synthetic polymers have been used to act as a scaffold for chondrocytes. The production of extracellular matrix proteins by chondrocytes has been studied. As tissue engineers, it is advantageous to explore the possibility of how altering biomaterial properties affect the signaling cascades by activation of receptors and transduction through the cytoplasm. This vital information will be able to aid in the future of engineering an appropriate biomaterial that can incorporate chondrocytes to act as a scaffold for articular cartilage.

Knee articular cartilage injuries in athletes present a therapeutic challenge and have been identified as an important cause of permanent disability because of the high mechanical joint stresses in athletes.

To determine whether microfracture treatment of knee articular cartilage injuries can return athletes to high-impact sports and to identify the factors that affect the ability to return to athletic activity.

Case series; Level of evidence, 4.

Thirty-two athletes who regularly participated in high-impact, pivoting sports before articular cartilage injury were treated with microfracture for single articular cartilage lesions of the knee. Functional outcome was prospectively evaluated with a minimum 2-year follow-up by subjective rating, activity-based outcome scores, and the ability for postoperative participation in high-impact, pivoting sports.

At last follow-up, 66% of athletes reported good or excellent results. Activity of daily living, Marx activity rating scale, and Tegner activity scores increased significantly after microfracture. After an initial improvement, score decreases were observed in 47% of athletes. Forty-four percent of athletes were able to regularly participate in high-impact, pivoting sports, 57% of these at the preoperative level. Return to high-impact sports was significantly higher in athletes with age <40 years, lesion size <200 mm(2), preoperative symptoms <12 months, and no prior surgical intervention.

Microfracture is an effective first-line treatment to return young athletes with short symptomatic intervals and small articular cartilage lesions of the knee back to high-impact athletics.

Limited information exists about the treatment of full-thickness articular cartilage lesions of the knee in adolescent athletes.

To evaluate the functional outcome and athletic activity after articular cartilage repair in the knees of adolescent athletes.

Case series; Level of evidence, 4.

Twenty adolescent athletes with full-thickness articular cartilage lesions of the knee were treated with autologous chondrocyte transplantation. Functional outcome was evaluated by subjective patient outcome rating, knee activity scores, and level of athletic participation.

At a mean of 47 months after autologous chondrocyte transplantation, 96% of adolescents reported good or excellent results with significant increases in postoperative Tegner activity scores and Lysholm scores. Ninety-six percent returned to high-impact sports and 60% to an athletic level equal or higher than that before knee injury. Return to preinjury sports correlated with shorter preoperative symptoms and a lower number of prior operations. All adolescents with preoperative symptoms < or =12 months returned to preinjury-level athletics, compared to 33% with preoperative intervals longer than 12 months.

Treatment of full-thickness articular injuries of the knee in adolescent athletes with autologous chondrocyte transplantation yields a high rate of functional success at a mean follow-up of 47 months. The rate of return to demanding athletic activities is higher in cases in which the preoperative duration of symptoms is 12 months or less.

Experimental data indicate that perichondrial grafting to restore articular cartilage defects will result in repair with hyaline-like cartilage. In contrast. debridement and drilling results in repair with fibro-cartilage. In this retrospective study the long-term clinical results of both procedures were compared to evaluate the theoretical benefit of repair with hyaline-like tissue.

From two independent studies patients were selected with a cartilage defect in their knee. The selection was performed using strict inclusion criteria published elsewhere [Bouwmeester et al. Int. Orthop. 21 (1997) 313]. The patients were treated with either a perichondrium transplantation (PT group, n = 14) or with an 'open' debridement and drilling procedure (DD group, n = 11). The results of both procedures after 10-11 years were evaluated using the Hospital for Special Surgery Knee Score (HSSS), X-ray examination, clinical examination and visual analogue scales (VAS) for pain during walking and at rest.

Both procedures resulted in a general improvement compared to the situation before the operation. After an average of 10 years in the PT group there were three failures, in the DD group none, success rates were 78% and 100%, respectively. When comparing the successful PT patients with the DD patients, there were no differences in HSSS and VAS data. Both groups showed an equal number of irregular operation surface sites on X-ray (PT 9/11 versus DD 8/10).

This study shows that clinically at 10 years follow-up no difference was observed between debridement and drilling and perichondrium transplantation for treatment of an isolated cartilage defect. This raises questions about ongoing research to develop methods in order to improve the results of debridement and drilling as therapy for an isolated cartilage defect in a young patient (< or = 40 years).

Several studies have reported enhanced repair of damaged cartilage following implantation of mesenchymal stem cells (MSCs) into full-thickness cartilage defects suggesting that the cells in the repair tissue were derived from the implant. However, it cannot be excluded that the enhanced tissue repair is derived from host cells recruited to the defect in response to the implant, rather than the re-population of the tissue by the implanted MSCs. Our objective was to study the short-term fate of fluorescently labeled MSCs after implantation into full-thickness cartilage defects in vivo. The fluorescent dye used in our studies did not affect MSC viability or their ability to undergo osteogenic and chondrogenic differentiation in vitro. MSC gelatin constructs were implanted into full-thickness cartilage defects in goats. These cells retained the dye and were detectable by histology and flow cytometry. At intervals spanning 2 weeks post-implantation we observed gradual loss of implanted cells in the defect as well as fragments of gelatin sponge containing labeled MSCs in deep marrow spaces indicating fragmentation, dislodgement and passive migration. Fluorescent labeling enabled us to determine whether the implanted cells were lost during early time points after implantation as well as their spatial orientation throughout the defect. By determining the fate of implanted cells, new biomaterials could be engineered to correct undesirable characteristics. Testing of new biomaterials in short-term in vivo models would provide faster optimization for cell retention needed for successful, long-term cartilage regeneration.

This study evaluated the constitutive insulin-like growth factor-I (IGF-I) gene expression pattern in spontaneously healing cartilage defects over the course of 16 weeks, and correlated the tissue morphology and matrix gene expression with IGF-I mRNA levels. Full-thickness 15 mm cartilage defects were debrided in the femoral trochlea of both femoropatellar joints of 8 horses and the healing defects examined 2, 4, 8, or 16 weeks after surgery. Samples were harvested for histologic assessment of tissue healing using H&E staining, toluidine blue histochemical reaction for proteoglycan deposition, and in situ hybridization and immunohistochemistry procedures to demonstrate collagen type II mRNA and protein expression. Total RNA was isolated for Northern analysis to measure cartilage matrix molecule expression, and for semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) to determine IGF-I gene expression patterns in healing cartilage defects. Full-thickness cartilage defects in horses were slow to heal compared to smaller lesions in similar locations in other animals. However, a progressive decline in tissue cellularity and vascularity, and increased tissue organization were observed on H&E stained specimens over the 16-week experiment. Evidence of early chondrogenic repair was detected through collagen type II in situ hybridization and immunohistochemistry. However, levels of collagen type II and aggrecan mRNA in lesions were not abundant on Northern analysis indicating incomplete chondrogenesis. IGF-I message expression followed a cyclic pattern with low levels at 2 weeks, followed by an increase at 4 and 8 weeks, and a subsequent decline at 16 weeks. There was no direct correlation between the stage of healing and cartilage matrix message expression, and the abundance of IGF-I mRNA in the healing lesions. In conclusion, this study demonstrated that the spontaneous healing of articular defects was accompanied by a temporal fluctuation in IGF-I gene expression which was discoordinate to the steady rise in expression of cartilage matrix molecules such as procollagen type II.

Within the bone marrow stroma there exists a subset of nonhematopoietic cells referred to as mesenchymal stem or mesenchymal progenitor cells. These cells can be ex vivo expanded and induced, either in vitro or in vivo, to terminally differentiate into osteoblasts, chondrocytes, adipocytes, tenocytes, myotubes, neural cells, and hematopoietic-supporting stroma. The multipotential of these cells, their easy isolation and culture, as well as their high ex vivo expansive potential make these cells an attractive therapeutic tool. In this work we will review the information dealing with the biology of mesenchymal progenitors as it has been revealed mainly by ex vivo studies performed with bone marrow-derived cells. The discussed topics include, among others, characteristics of mesenchymal progenitors, evidence for the existence of a vast repertoire of uncommitted and committed progenitors both in the bone marrow and in mesenchymal tissues, a diagram for their proliferative hierarchy, and comments on mobilization, microenvironment, and clinical use of mesenchymal progenitors. Despite the enormous data available at molecular and cellular levels, it is evident that a number of fundamental questions still need to be resolved before mesenchymal progenitors can be used for safe and effective clinical applications in the context of both cell and gene therapies.

It is well established that a full-thickness articular cartilage defect is repaired with a fibrocartilaginous tissue of which cells are derived from undifferentiated mesenchymal stem cells in the bone marrow. To characterize the repair tissue immunohistochemically, full-thickness defects were created in rabbit knee joints, and the repair tissues immunostained at 3, 6, and 12 weeks after surgery. Well characterized polyclonal antibody against carboxyterminal type II procollagen peptide (pCOL-II-C) and monoclonal antibody against type II collagen were used to evaluate the repair tissue with regard to the metabolism of type II collagen. Immunohistochemistry revealed that pCOL-II-C was localized in or around most of the repair cells obtained at 3 and 6 weeks after surgery, while type II collagen distributed mainly in the pericellular matrix of metaplastic round-shaped repair cells. The results suggest that the repair cells taken at the early stage of the repair process of the defect could originally have more activity of type II collagen synthesis.