JOURNAL ARTICLE

[Effects of bone marrow mesenchymal stem cells with acellular muscle bioscaffolds on repair of acute hemi-transection spinal cord injury in rats]

Xiangke Wei, Yimin Wen, Tao Zhang, Han Li
Chinese Journal of Reparative and Reconstructive Surgery 2012, 26 (11): 1362-8
23230674

OBJECTIVE: To investigate the effects of allogenic transplantation of acellular muscle bioscaffolds (AMBS) seeded with bone marrow mesenchymal stem cells (BMSCs) on the repair of acute hemi-transection spinal cord injury (SCI) in rats.

METHODS: AMBS were prepared by reformed chemical approach and sterilized by compound cold sterilization; BMSCs were harvested by density gradient centrifugation and cultured with adherent method. The 3rd generation BMSCs labeled by Hoechst 33342 were injected into AMBS to construct the BMSCs-AMBS composite scaffolds; the biocompatibility was observed under scanning electron microscope (SEM) and fluorescence microscope in vitro at 14 days. Forty-eight adult female Sprague Dawley rats were used to build SCI model by hemi-transecting at T(9-11) level, then randomly divided into 4 groups (n=12). Defects were repaired with BMSCs-AMBS composite scaffolds, BMSCs, and AMBS in groups A, B, and C, respectively; group D was blank control by injecting PBS. At 1, 2, 3, and 4 weeks after surgery, the functional recovery of the hind limbs was evaluated by the Basso-Beattie-Bresnahan (BBB) locomotor rating score. At 4 weeks after surgery, HE staining and immunofluorescent assay were adopted.

RESULTS: Masson staining and HE staining showed that AMBS was mainly of the collagen fibers in parallel arrange, without muscle fibers. After 14 days of BMSCs and AMBS co-culture, a large number of survival BMSCs labeled by Hoechst 33342 were seen under fluorescence microscope; SEM showed that BMSCs grew and attached to the inner surfaces of AMBS. At 2-4 weeks, the BBB score in group A was significantly higher than that in groups B, C, and D (P < 0.05), and it was significantly lower in group D than in the other 3 groups (P < 0.05); at 4 weeks, the BBB score in group B was significantly higher than that in group C (t=10.352, P=0.000). HE staining revealed that the area of spinal cord cavity after SCI was markedly smaller in group A than in the other 3 groups; immunofluorescent assay showed that more neurofilament 200 positive fibers and Nestin positive cells were detected in group A than in groups B, C, and D, but glial fibrillary acidic protein (GFAP) positive cells significantly decreased. The integral absorbance (IA) values of GFAP were 733.01 +/- 202.04, 926.42 +/- 59.46, 1 069.37 +/- 33.42, and 1 469.46 +/- 160.53 in groups A, B, C, and D, respectively; the IA value of group A was significantly lower than that of groups B, C, and D (P < 0.05), and it was significantly higher in group D than in groups A, B, and C (P < 0.05).

CONCLUSION: With relatively regular internal structures and good biocompatibility, AMBS can inhibit glial scar and enhance the survival, migration, and differentiation of BMSCs, so AMBS is the ideal nature vector for cell transplantation. Co-transplantation of AMBS and BMSCs has synergistic effect in treating SCI, it can promote rat motor function recovery.

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