Among these cytokines, bFGF is one of the most potent mitogens for therapeutic angiogenesis selleck chemical [18]. As chronic ischemic hindlimb needs long-term treatment, a sustained drug delivery system is urgently needed. The duration of exposure to bFGF may be a critical determinant of formation of stable neovasculature [19].Biosynthetic matrices with bioactive and bioresorbable characteristics are becoming more important than ever before [20, 21]. Future biosynthetic matrices should possess both bioactive and bioresorbable properties to facilitate tissue regeneration and enhance the replacement of the matrix itself [22, 23]. Our previous report confirmed that gelatin microspheres loaded with bFGF improved capillary formation in the lower extremity compared with bFGF alone [11].
Therefore in the present study, we used another common biodegradable material, that is, collagen.According to previous studies, the collagen biomaterial has prominent properties: firstly, the pore and porosity of the collagen matrix could be controlled [24]; secondly, cytokines could easily be impregnated into the collagen matrix without affecting its bioactivity [25]; thirdly, a stable and long-term sustained release of cytokines from the collagen matrix could be achieved [25]. Another reason for using a collagen matrix is based on our prior work in which we demonstrated the utility of collagen biomaterial for cell attachment and proliferation [26]. Moreover, collagen scaffold seeded with bone marrow-derived MSCs was a stronger angiogenic inducer than MSCs alone [27].
Collagen can induce proangiogenesis and provide long-term activation of gelatinase A, which is likely to be relevant to endothelial cell invasion during angiogenesis [28]. Our results were consistent with these studies.The bFGF was released from the CM over a period of time and triggered therapeutically significant angiogenesis, including improved hindlimb perfusion and the formation of capillaries and mature blood vessels. These results suggest that angiogenesis depends on the sustained release of bFGF.The study lasted for 4 weeks in order to decrease the formation of leaky vessels and facilitate the formation of pericytes or smooth muscle cells indicating the stable vasculature around endothelial cells [29]. In the bFGF-CM group, the oxygen saturation, capillary growth, and mature vessel formation were significantly more than those in the CM, bFGF, and ischemic groups.
The percentages of the mature vessels/the total vessels in bFGF-CM, CM, bFGF, and ischemic groups were 24.8%, 29.8%, 33.3%, and 23.7%, respectively. There was more capillaries formation than mature vessels formation at 4 weeks. We speculated that if we prolonged the research, more capillaries AV-951 would become mature vessels due to the bFGF sustained releasing profile.
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