Degraded RNA from Human Anterior Cruciate Ligaments Yields Valid Gene Expression Profiles.

Correlating gene expression patterns with biomechanical properties of connective tissues provides insights into the molecular processes underlying the tissue growth and repair. Cadaveric specimens such as human knees are widely considered suitable for biomechanical studies, but their usefulness for gene expression experiments is potentially limited by the unavoidable, nuclease-mediated degradation of RNA. Here, we tested whether valid gene expression profiles can be obtained using degraded RNA from human anterior cruciate ligaments (ACLs). Human ACL RNA (N = 6) degraded in vitro by limited ribonuclease digestion resemble highly degraded RNA isolated from cadaveric tissue. PCR threshold cycle (Ct) values for 90 transcripts (84 extracellular matrix, 6 housekeeping) in degraded RNAs variably ranged higher than values obtained from their corresponding non-degraded RNAs, reflecting both the expected loss of target templates in the degraded preparations as well as differences in the extent of degradation. Relative Ct values obtained for mRNAs in degraded preparations strongly correlated with the corresponding levels in non-degraded RNA, both for each ACL as well as for the pooled results from all six ACLs. Nuclease-mediated degradation produced similar, strongly correlated losses of housekeeping and non-housekeeping gene mRNAs. RNA degraded in situ yielded comparable results, confirming that in vitro digestion effectively modeled degradation by endogenous ribonucleases in frozen and thawed ACL. We conclude that, contrary to conventional wisdom, PCR-based expression analyses can yield valid mRNA profiles even from RNA preparations that are more than 90% degraded, such as those obtained from connective tissues subjected to biomechanical studies. Furthermore, legitimate quantitative comparisons between variably degraded tissues can be made by normalizing data to appropriate housekeeping transcripts.

Influence of a platelet concentrate on prosthetic bone ingrowth in a rabbit model.

Recent studies have shown that an increase in bone ingrowth by addition of osteogenic growth factors can reduce micro motion and gross implant motion and contribute to joint implant stability through osseointegration. Platelet-rich plasma (PRP) has the potential to provide growth factors that may be conducive to osteointegration at the bone-implant interface. This study analyzed the influence of PRP on bone ingrowth upon a beaded metal implant in distal femurs of 22 rabbits. Rabbit limbs were randomly assigned to receive an implant plus PRP or plain implant. Half of the specimens were randomly assigned to a 2-week group (n = 20) or a 5-week group (n = 20). Histologic and histomorphometric comparison between implant alone and implant plus PRP, at 2 and 5 weeks, was performed. In both the 2- and 5-week comparisons, there was no statistical difference (p > .05) in bone ingrowth between the control and PRP group, despite a slight increase in trabecular bone growth in PRP groups. This study suggests that PRP is not a major contributing factor to bone ingrowth at the bone-implant interface. This supports growing evidence in the literature that PRP can lead to variable bone growth stimulation in vivo.

Osteochondral autograft transplantation in the porcine knee.

BACKGROUND: Knee articular cartilage defects are not an uncommon problem. Because articular cartilage is limited in its ability to heal, these defects are difficult to manage. HYPOTHESIS: Osteochondral autografts will provide less of a cavitary defect and more viable hyaline articular cartilage than will control knees. STUDY DESIGN: Controlled laboratory study. METHODS: Osteochondral autografts were grossly and microscopically evaluated in the porcine knee and compared with a control at 6 weeks, 3 months, and 6 months. In 18 porcine specimens, a 1-stage surgical procedure was performed to harvest an osteochondral graft from a nonweightbearing articular cartilage surface, and the graft was transplanted into a defect created in the weight-bearing region of the medial femoral condyle. In the opposite control knee, a similar defect was created in the medial femoral condyle; an osteochondral transplant was not performed. Six pigs each were sacrificed at 6 weeks, 3 months, and 6 months. RESULTS: Gross inspection of the control knees showed a cavitary defect. The defect grossly decreased in size with fibrous ingrowth seen on microscopic analysis. An increasing amount of fibrous tissue and fibrocartilage was present at the 3 time periods. Gross inspection of the graft knee showed a healed osteochondral plug with no obvious displacement, cavitary defects, or surrounding necrotic tissue at each time interval. Microscopic analysis revealed the graft knee contained viable hyaline cartilage and healed viable subchondral bone. At all time intervals, 75% to 100% of the hyaline cartilage was viable in all specimens. In 6-month specimens, bridging cartilage at the autograft-host junction was incomplete in 50%, partial in 33%, and complete in 17%. CONCLUSION: Osteochondral autografts in the porcine knee resulted in viable hyaline cartilage for up to 6 months; there was inconsistent bridging hyaline cartilage at the periphery. Grafts appeared to heal into existing subchondral bone without displacement or evidence of necrosis. CLINICAL RELEVANCE: This type of osteochondral transplant can be used as a reliable reconstructive alternative for osteochondral defects.

Case reports: malignant transformation of aneurysmal bone cysts.

An aneurysmal bone cyst is an uncommon benign primary bone tumor. Careful intralesional curettage through a wide cortical window in addition to cauterization with or without adjuvant therapy (phenol or hydrogen peroxide) and bone grafting or cementation is the preferred surgical treatment. Adjuvant or primary radiation of an aneurysmal bone cyst rarely is used because of its association with malignant transformation of the lesion. Several cases of late malignant transformation of primary aneurysmal bone cysts without adjuvant radiation have been reported. We provide additional documentation of two primary aneurysmal bone cysts treated surgically with careful intralesional curettage through a wide cortical window and allograft bone grafting without adjuvant radiation. At 5.5 years and 12 years after treatment, a telangiectatic osteosarcoma and a fibroblastic osteosarcoma, respectively, were identified in the site of the original lesions. Not only should aneurysmal bone cysts be evaluated carefully through histologic examination at presentation, patients also should be counseled regarding possible recurrence and the need for routine followups, especially if symptoms change.