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. 2019 Nov 22;20(23):5859. doi: 10.3390/ijms20235859

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© 2019 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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Forced expression of miR-29a or transplantation of human umbilical cord Wharton’s jelly (WJ)-derived mesenchymal stem cells (uMSCs) successfully restored the blood perfusion in the anesthetized C57BL/6 mice post BPVC-injury (n = 6/each group). Representative laser Doppler perfusion images of both left hind (LH) and right hind (RH) limbs. After 7 days of BPVC injury on the LH limb, impaired blood perfusion was shown in ((B); LH site), while compared to the saline injected LH limb Sham group (A) or its contralateral control RH limb ((B); RH site). Injection of Pre miR-29a notably rescued the BPVC-suppressed blood perfusion in LH limb ((C); LH site) whereas the injection of scramble NC (negative control of miR-29a) into the RH limb did not have any significant restoration on perfusion recovery ((C); RH site). (D) Injection of miR-29a AS (inhibitor of miR-29a) into BPVC-induced ischemic LH limb aggravated the impairment of blood perfusion. (E) Transplantation of uMSCs into BPVC-injured LH limbs considerably restored the perfusion while compared to the BPVC-injured LH limb in ((B); LH site). Of note, transplantation of uMSCs together with Pre miR-29a conspicuously renovated the BPVC-induced LH limb ischemic perfusion and shown in (F). (G) Quantitative RT-PCR analysis confirmed that Pre miR-29a injection group strongly displayed miR-29a transcripts in gastrocnemius muscles collected from ((C); LH site) than those in the scramble NC treatment group collect from ((C); RH site). In contrast, the level of miR-29a was evidently suppressed in the miR-29a AS treatment groups shown in ((D); LH site) than those in scramble NC group shown in ((D); RH site). (H) Histograms showed the quantification of blood flow perfusion unit (PU) per field which underwent saline-or BPVC injection, Pre miR-29a or miR-29a AS administration, transplantation of uMSCs, and combination of Pre miR-29a and the uMSCs treatments post BPVC injury. Data were given as means ± SD; *** p < 0.001 for the BPVC group vs. the Sham group or the BPVC + scramble NC group vs. Sham + scramble NC group; # p < 0.05, ## p < 0.01, ### p < 0.001 for the BPVC group vs. the Sham group; * p < 0.05 for the BPVC + Pre miR-29a group, BPVC + the miR-29a AS group, the BPVC + uMSC group, the BPVC + uMSC + Pre miR-29a group vs. the BPVC group.