Our data show that the application of a PRP fibrin membrane around the neurorraphy improves the nerve regeneration process in a rat sciatic nerve model. The use of PRP as a suturable membrane could perform an action not only as a source of bioactive proteins but also as a nerve guide to hold the scar reaction and thus improve axonal regeneration.
Stem cells from extra-embryonic sources can be obtained by non-invasive procedures. We have standardized a method for the expansion of equine umbilical cord-derived matrix cells (EUCMCs) for potential therapy. EUCMCs were isolated from the umbilical cord of five mares immediately after delivery. For expansion, cells were grown in alpha-MEM and MSCBM. Moreover, to measure the effect of growth factor supplementation, epidermal growth factor (EGF) was added to alpha-MEM. alpha-MEM and MSCBM media performed similarly in terms of population doubling and CFU number value. EGF supplementation of alpha-MEM determined a significant increase of the population doubling value. EGF supplementation did not affect the adipogenic and chondrogenic differentiation while bone nodule sizes an increased with the osteogenic protocol. Both alpha-MEM and MSCBM can be used to cultivate EUCMCs. alpha-MEM supplemented with EGF might represent an advantage for EUCMCs expansion. The results could be useful in choosing the culture medium since alpha-MEM is more cost-effective than MSCBM.
Background -: Rabbits provide an excellent model for many animal and human diseases, such as cardiovascular diseases, for the development of new vaccines in wound healing management and in the field of tissue engineering of tendon, cartilage, bone and skin.
One of the most challenging requirements of a successful bone tissue engineering approach is the development of scaffolds specifically tailored to individual tissue defects. Besides materials chemistry, well-defined scaffold’s structural features at the micro- and macro-levels are needed for optimal bone in-growth. In this study, polymeric fibrous scaffolds with a controlled internal network of pores and modelled on the anatomical shape and dimensions of a critical size bone defect in a rabbit’s radius model were developed by employing a computer-aided wet-spinning technique. The tailored scaffolds made of star poly(ε-caprolactone) or star poly(ε-caprolactone)–hydroxyapatite composite material were implanted into 20-mm segmental defects created in radial diaphysis of New Zealand white rabbits. Bone regeneration and tissue response were assessed by X-rays and histological analysis at 4, 8 and 12 weeks after surgery. No signs of macroscopic and microscopic inflammatory reactions were detected, and the developed scaffolds showed a good ability to support and promote the bone regeneration process. However, no significant differences in osteoconductivity were observed between star poly(ε-caprolactone) and star poly(ε-caprolactone)–hydroxyapatite scaffolds. Long-term study on implanted star poly(ε-caprolactone) scaffolds confirmed the presence of signs of bone regeneration and remodelling, particularly evident at 24 weeks.
The amnion is a particular tissue whose cells show features of multipotent stem cells proposed for use in cellular therapy and regenerative medicine. From equine amnion collected after the foal birth we have isolated MSCs (mesenchymal stem cells), namely EAMSCs (equine amnion mesenchymal stem cells), from the mesoblastic layer. The cells were grown in α-MEM (α-modified minimum essential medium) and the effect of EGF (epidermal growth factor) supplementation was evaluated. To assess the growth kinetic of EAMSCs we have taken into account some parameters [PD (population doubling), fold increase and DT (doubling time)]. The differentiation in chondrogenic, adipogenic and osteogenic types of cells and their epitope expression by a cytofluorimetric study have been reported. EGF supplementation of the culture medium resulted in a significant increase in PD growth parameter and in the formation of bone nodules for the osteogenic differentiation. By immunohistochemistry the amnion tissue shows a positivity for the c-Kit (cluster tyrosine-protein kinase), CD105 and Oct-4 (octamer-binding transcription factor 4) antigens that confirmed the presence of MSCs with embryonic phenotype.
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