Fig 1.
Maximal isometric tension of soleus muscle after BMMC treatment.
(A) Maximal isometric force in normal (non-injured), BMMC and vehicle-treated groups 14 days after injury. (B) Same as in (A) except that animals were analyzed 28 days after injury. Normal group values are from contralateral non-injured muscle. Values are means ± SD. *** p<0.001 * p<0.05. BMMC (14d n = 6 and 28d n = 9) and vehicle-treated rats (14d n = 6 and 28d n = 7)
Fig 2.
Histological sections stained by heamatoxylin & eosin.
(A1) H&E staining image of a rat non-injured soleus muscle 28 days after surgery; (A2) same as in A1 except that vehicle-treated muscle is shown; (A3) same as in A1 but BMMC-treated animal is presented; (A2, A3 and A4) Details of A1, A2 and A3 animals are shown in greater magnification, respectively; (A3) Picrossírius staining image of a rat non-injured soleus muscle 28 days after surgery;; (B3) same as in A3 except that vehicle-treated muscle is shown; (C3) same as in A3 but BMMC-treated animal is presented and (A4, B4 and C4) Details of A3, B3 and C3 animals are shown in greater magnification, respectively; Histological analysis reveals that both groups submitted to muscle injury present wide spread collagen deposition (A3-C4) independent of treatment. Note the central aligned disposition of the muscle fiber nuclei indicating recent muscle fiber regeneration (arrows in B2). Bars correspond to 300 μm in A1-C1 and A3-C3 and correspond to 50μm in A2-C2 and A4-C4.
Table 1.
Quantification of collagen content.
Fig 3.
Centronucleated muscle fiber and cross-sectional area determination 28 days after muscle injury.
(A) Number of centronucleated muscle fibers per 103 fibers in normal (non-injured), vehicle- and BMMC-treated groups; (B) muscle fiber cross-sectional area in same groups as A; (C) Cross-sectional image a normal soleus muscle stained with H&E; (D) Same as in C but vehicle muscle is shown and (E) same as in C but BMMC soleus muscle is presented. Normal group values are from contralateral non-injured muscle. Values are means ± SD. *** p<0.001 * p<0.05. BMMC (n = 4) and vehicle-treated rats (n = 3). Centronucleated muscle fibers are shown by arrow reads.
Fig 4.
Skeletal muscle troponin immunolocalization in soleus muscle treated with BMMC 28 days after repeated injuries.
(A) and (D) Differential interference contrast images; (B) and (E) overlay images of Hoestch (in blue) and troponin I (green); (C) and (F) troponin I staining alone. Green fluorescence indicates typical striated skeletal muscle labelling pattern while nuclei are shown by blue Hoescht staining. Arrows denote nuclei position. Bars correspond to 20μm.
Fig 5.
Immunolocalization of smooth muscle myosin as vessel marker in soleus muscle treated with BMMC 28 days after repeated injuries.
A) and (D) Differential interference contrast images; (B) and (E) overlay images of Hoestch (in blue) and smooth muscle myosin (green) staining; (C) and (F) smooth muscle myosin alone. Green fluorescence indicates typical vessel shape labelling pattern while nuclei are shown by blue Hoescht staining. Arrows denote nuclei position. Bars correspond to 20μm.