Validity of radiographic evaluations of bone formation in a rat calvaria osteotomy defect model.

OBJECTIVE: The objective of this study was to evaluate the validity of radiographic evaluations of bone formation in a critical-size rat calvaria osteotomy defect model. METHODS: Bilateral, critical-size ( [symbol in text] 6 mm) calvaria osteotomy defects in 30 adult Sprague-Dawley rats treated with a rat platelet-rich plasma preparation or control treatments were evaluated by radiographic and histometric measures following a 4- or 8-week healing interval. Standardized radiographic images of the rat calvaria gross specimens were used to assess bone formation within the defect sites by visual evaluation of the grey scale by three masked examiners. The most central portion of each defect site was subject to histometric analysis using a PC-based image analysis system. Kappa statistics and percentage agreement between the radiographic and histometric analysis were estimated. RESULTS: Radiographic evaluations of bone formation are associated with significant weaknesses poorly representing actual healing events; kappa statistics (0.17) denoting slight agreement beyond chance. Perfect agreement between the histologic and radiographic analysis for defect sites showing complete and partial histologic bone fill was achieved 63% and 50% of the time, respectively. Agreement reached only 20% for sites with no/limited bone fill. When no/limited and partial bone fill occurred, the radiographic analysis tended to overestimate bone fill and underestimate bone fill when complete closure of the defect sites was observed in the histologic analysis. CONCLUSION: Low accuracy was observed when radiographic evaluations were employed in identifying and characterizing bone fill in the rat calvaria osteotomy defects. Assessment of bone healing in animal models aiming at treatment recommendations for clinical application must not solely be based on radiographic analysis, but should be confirmed using histologic observations.

Analysis of rat calvaria defects implanted with a platelet-rich plasma preparation: radiographic observations.

BACKGROUND: Platelet-rich plasma (PRP) harbors growth factors identified in bone. It has been suggested that these factors enhance osteogenesis. The objective of this study was to conduct a radiographic evaluation on local bone formation following surgical implantation of a PRP preparation using a critical-size rat calvaria defect model. METHODS: Thirty 22-week-old male Sprague-Dawley rats were used. The PRP preparation was obtained from 10 ml of whole blood drawn from one age-matched donor rat. The preparation was processed by gradient density centrifugation and stored at -80 degrees C until use. Using aseptic techniques, the PRP preparation soak-loaded onto an absorbable collagen sponge (ACS) carrier or ACS alone was surgically implanted into contralateral critical-size 6 mm rat calvaria osteotomies in 18 animals. Twelve animals received ACS alone versus sham surgery in contralateral defects. Animals were sacrificed at 4 and 8 weeks when biopsies were collected and radiographs were obtained using a standardized protocol. Three masked examiners independently evaluated the radiographic images of the defect sites. Examiner reproducibility was examined by repeat evaluation of all defect sites (r=0.6; P <0.0001). RESULTS: The animals were maintained without adverse events. Defect sites in two animals receiving ACS versus sham surgery (4-week healing interval) were not evaluated due to specimen damage. Seventy-five percent of the sites (PRP/ACS or ACS) exhibited partial closure at 4 weeks; one site (ACS) exhibited full closure without significant differences between protocols (P=0.1797). Fifty percent of the sites receiving PRP/ACS exhibited full closure and 20% partial closure at 8 weeks versus 20% and 80%, respectively, for the ACS control (P=0.7532). There were no noteworthy differences between sites receiving ACS versus sham surgery at 4 or 8 weeks. CONCLUSION: The results suggest that the PRP preparation does not have a significant effect on osteogenesis.

Analysis of rat calvaria defects implanted with a platelet-rich plasma preparation: histologic and histometric observations.

OBJECTIVES: It has been suggested that degranulating platelet alpha-granules release growth factors having a potential to modulate bone formation. The objective of this study was to evaluate the osteoconductive potential of a platelet-rich plasma (PRP) preparation. METHODS: Thirty adult male Sprague-Dawley rats were used. The PRP preparation was obtained from 10 ml of whole blood drawn from one age-matched donor rat. The preparation was processed by gradient density centrifugation and stored at -80 degrees C until use. Using aseptic techniques, the PRP preparation soak loaded onto an absorbable collagen sponge (ACS) or ACS alone was surgically implanted into contralateral critical size 6-mm calvaria osteotomies in 18 animals. Twelve animals received ACS versus sham surgery in contralateral defects. Animals were sacrificed at 4 and 8 weeks when biopsies were collected for histologic and histometric analysis. RESULTS: The animals were maintained without adverse events. Bone formation was highly variable in sites receiving PRP and control treatments. Defect bone fill at 4 weeks averaged (+/-SD) 28.8+/-27.4% (PRP/ACS) versus 39.1+/-24.4% (ACS; p=0.2626) and 62.0+/-20.0% (ACS) versus 71.6+/-32.2% (sham surgery; p=0.1088), and at 8 weeks 81.0+/-12.9% (PRP/ACS) versus 64.5+/-28.1% (ACS; p=0.2626) and 75.6+/-34.1% (ACS) versus 74.1+/-24.2% (sham surgery; p=0.7353). Remnants of the ACS biomaterial were observed at both 4 and 8 weeks in sites implanted with PRP/ACS or ACS. CONCLUSIONS: The results suggest that the PRP preparation has a limited potential to promote local bone formation.