Efficacy of intraoperatively prepared cell-based constructs for bone regeneration.

BACKGROUND: Conventional cell-based bone regeneration suffers from the major disadvantage of limited cell supply, time-consuming in vitro expansion cultures, and limited patient-friendliness related to cell isolation and multiple visits to the clinic. Here, we utilized an alternative concept using "easy access cells" that can be obtained in an intraoperative manner to prepare cell-based constructs. METHODS: We used stromal vascular fraction (SVF) from human adipose tissue and human monocytes for intraoperative preparation of bone constructs. Conventional constructs grafted with expanded human adipose tissue mesenchymal stem cells (ADMSCs) derived from the same donor were set as positive controls. Additionally, we combined both cell types either or not with monocytes. The cellular interaction of human SVF and ADMSCs with human monocytes was evaluated in vitro. The feasibility and bone-regenerative capacity of intraoperative constructs were determined histologically and histomorphometrically in a rat femoral condyle bone defect model. RESULTS: SVF displayed equal in vitro osteogenic differentiation compared to donor-matched expanded ADMSCs, which for both was significantly enhanced upon co-culture with monocytes. Moreover, SVF and ADMSCs displayed different immunoregulatory effects on monocytes/macrophages. Upon implantation in rat femoral bone defects, SVF constructs demonstrated superior bone formation compared to ADMSC constructs and cell-free controls; no effects of monocyte addition were observed. CONCLUSION: In conclusion, we here demonstrate the feasibility of intraoperative SVF construct preparation and superior bone-regenerative capacity thereof compared to donor-matched ADMSC constructs. The superiority of SVF constructs was found to be linked to the distinct differences between immunoregulatory effects of SVF and ADMSCs.

Biomaterial Property Effects on Platelets and Macrophages: An in Vitro Study.

The purpose of this study was to evaluate the effects of surface properties of bone implants coated with hydroxyapatite (HA) and ß-tricalcium phosphate (ß-TCP) on platelets and macrophages upon implant installation and compare them to grit-blasted Ti and Thermanox used as a control. Surface properties were characterized using scanning electron microscopy, profilometry, crystallography, Fourier transform infrared spectroscopy, and coating stability. For platelets, platelet adherence and morphology were assessed. For macrophages, morphology, proliferation, and polarization were evaluated. Surface characterization showed similar roughness of ∼2.5 µm for grit-blasted Ti discs, both with and without coating. Coating stability assessment showed substantial dissolution of HA and ß-TCP coatings. Platelet adherence was significantly higher for grit-blasted Ti, Ti-HA, and Ti-ß-TCP coatings compared to that of cell culture control Thermanox. Macrophage cultures revealed a decreased proliferation on both HA and ß-TCP coated discs compared to both Thermanox and grit-blasted Ti. In contrast, secretion of pro-inflammatory cytokine TNF-α and anti-inflammatory cytokine TGF-ß were marginal for grit-blasted Ti and Thermanox, while a coating-dependent increased secretion of pro- and anti-inflammatory cytokines was observed for HA and ß-TCP coatings. The results demonstrated a significantly upregulated pro-inflammatory and anti-inflammatory cytokine secretion and marker gene expression of macrophages on HA and ß-TCP coatings. Furthermore, HA induced an earlier M1 macrophage polarization but more M2 phenotype potency than ß-TCP. In conclusion, our data showed that material surface affects the behaviors of first cell types attached to implants. Due to the demonstrated crucial roles of platelets and macrophages in bone healing and implant integration, this information will greatly aid the design of metallic implants for a higher rate of success in patients.

Porous poly (D,L-lactide-co-glycolide) acid/biosilicate® composite scaffolds for bone tissue engineering.

This study evaluated the effects of the Biosilicate® and poly (D,L-lactic-co-glycolic) acid composites on bone repair in a tibial bone defect model in rats by means of using histological evaluation (histopathological and morphometric analysis) and gene expression analysis. Eighty male Wistar rats (12 weeks old, weighing ±300 g) were randomly divided into two groups: Biosilicate® group (BG) and Biosilicate® /PLGA group (BG/PLGA). Each group was euthanized at 3, 7, 14, and 21 days after surgery (n = 10 animals per time point). The main findings showed that the incorporation of PLGA into BG had a significant effect on the morphological structure of the material, accelerating mass loss, decreasing the pH and increasing the calcium release. Furthermore, histologic analysis revealed that the BG/PLGA showed increased material degradation, accompanied by higher bone formation compared to BG, after 21 days of implantation. In addition, qRT-PCR analysis showed that BG/PLGA induced an upregulation of the osteogenic genes related to BMP4, Runx2, ALP, and OC. These results show that the present BG/PLGA composite may be used as a bone graft for inducing bone repair. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 63-71, 2017.

Effect of low-level laser therapy (808 nm) on skeletal muscle after endurance exercise training in rats.

BACKGROUND: Low-level laser therapy (LLLT) has been demonstrated to be effective in optimizing skeletal muscle performance in animal experiments and in clinical trials. However, little is known about the effects of LLLT on muscle recovery after endurance training. OBJECTIVE: This study evaluates the effects of low-level laser therapy (LLLT) applied after an endurance training protocol on biochemical markers and morphology of skeletal muscle in rats. METHOD: Wistar rats were divided into control group (CG), trained group (TG), and trained and laser irradiated group (TLG). The endurance training was performed on a treadmill, 1 h/day, 5 days/wk, for 8 wk at 60% of the maximal speed reached during the maximal effort test (Tmax) and laser irradiation was applied after training. RESULTS: Both trained groups showed significant increase in speed compared to the CG. The TLG demonstrated a significantly reduced lactate level, increased tibialis anterior (TA) fiber cross-section area, and decreased TA fiber density. Myogenin expression was higher in soleus and TA muscles in both trained groups. In addition, LLLT produced myogenin downregulation in the TA muscle of trained animals. CONCLUSION: These results suggest that LLLT could be an effective therapeutic approach for stimulating recovery during an endurance exercise protocol.

Effect of low-level laser therapy (808 nm) on skeletal muscle after endurance exercise training in rats

BACKGROUND: Low-level laser therapy (LLLT) has been demonstrated to be effective in optimizing skeletal muscle performance in animal experiments and in clinical trials. However, little is known about the effects of LLLT on muscle recovery after endurance training. OBJECTIVE: This study evaluates the effects of low-level laser therapy (LLLT) applied after an endurance training protocol on biochemical markers and morphology of skeletal muscle in rats. METHOD: Wistar rats were divided into control group (CG), trained group (TG), and trained and laser irradiated group (TLG). The endurance training was performed on a treadmill, 1 h/day, 5 days/wk, for 8 wk at 60% of the maximal speed reached during the maximal effort test (Tmax) and laser irradiation was applied after training. RESULTS: Both trained groups showed significant increase in speed compared to the CG. The TLG demonstrated a significantly reduced lactate level, increased tibialis anterior (TA) fiber cross-section area, and decreased TA fiber density. Myogenin expression was higher in soleus and TA muscles in both trained groups. In addition, LLLT produced myogenin downregulation in the TA muscle of trained animals. CONCLUSION: These results suggest that LLLT could be an effective therapeutic approach for stimulating recovery during an endurance exercise protocol.