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1.
J Cell Biochem ; 113(7): 2330-45, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22573552

RESUMO

The expression of inflammatory cytokines and growth factors in surgically repaired lacerated muscles over a 12-week recovery phase was investigated. We hypothesized that these expression levels are influenced by both neural and muscular damage within lacerated muscles. Microarrays were confirmed with reverse transcription-polymerase chain reaction assays and histology of biopsies at the lesion of three simulated lacerated muscle models in 130 adult rats. The lacerated medial gastrocnemius with the main intramuscular nerve branch either cut (DN), crushed but leaving an intact nerve sheath (RN); or preserved intact (PN) were compared. At 4 weeks, DN had a higher number of interleukins up-regulated. DN and RN also had a set of Bmp genes significantly expressed between 2 and 8 weeks (P ≤ 0.05). By 12 weeks, DN had a poorer and slower myogenic recovery and greater fibrosis formation correlating with an up-regulation of the Tgf-ß gene family. DN also showed poorer re-innervation with higher mRNA expression levels of nerve growth factor (Ngf) and brain-derived neurotrophin growth factor (Bdnf) over RN and PN. This study demonstrates that the inflammatory response over 12 weeks in lacerated muscles may be directed by the type of intramuscular nerve damage, which can influence the recovery at the lesion site. Inflammatory-related genes associated to the type of intramuscular nerve damage include Gas-6, Artemin, Fgf10, Gdf8, Cntf, Lif, and Igf-2. qPCR also found up-regulation of Bdnf (1-week), neurotrophin-3 (2w), Lif (4w), and Ngf (4w, 8w) mRNA expressions in DN, making them possible candidates for therapeutic treatment to arrest the poor recovery in muscle lacerations (250).


Assuntos
Citocinas/biossíntese , Músculo Esquelético/lesões , Músculo Esquelético/inervação , Fator de Crescimento Neural/biossíntese , Regeneração Nervosa , Fator de Crescimento Transformador beta/biossíntese , Animais , Fator Neurotrófico Derivado do Encéfalo/biossíntese , Citocinas/metabolismo , Fibrose/patologia , Inflamação/imunologia , Fator Inibidor de Leucemia/biossíntese , Músculo Esquelético/patologia , Músculo Esquelético/cirurgia , Compressão Nervosa , Fator de Crescimento Neural/genética , Neurotrofina 3/biossíntese , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Fator de Crescimento Transformador beta/genética
2.
J Orthop Res ; 28(6): 732-8, 2010 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-20041489

RESUMO

Lacerated skeletal muscles often do not recover full function after repair. Denervated muscles with altered myosin heavy chain isoform (MHC) profiles are known to result in functional impairment. We studied the functional recovery of lacerated muscles, assessing MHC profile changes in association to the involvement of the intramuscular nerve (IM). We tested three lacerated models using the rabbit's medial gastrocnemius where the IM was either cut (NNR), repaired (NR), or preserved intact (NP). Muscles were assessed 7 months after repair for muscle atrophy, isometric contraction (by electrical stimulation), and fibrosis formation at the lesion site. Changes in myofibrillar actomyosin adenosine triphosphatase activity, MHC profile, regenerating myofibers and reinnervation were assessed by Western blot, histology, or immunohistology. Lacerated muscles with a repaired (NR) or an intact (NP) IM showed good recovery, with no significant changes in the MHC profile. Muscles where the IM was not repaired (NNR) resulted in significant scar area at the lesion site (p < 0.05), muscle atrophy (67%, p < 0.05) and loss in contractile properties (63% of the uninjured side, p < 0.05). At 7 months, all muscles were reinnervated. However, the NNR had an inappropriate (polyneural) and poorly distributed reinnervation, the presence of regenerating myofibers, and demonstrated a fast-to-slow MHC transition (71%:29% to 44%:56%, ANOVA, p = 0.018). This was associated to the cut IM when the NNR muscle was lacerated. Poor reinnervation in lacerated skeletal muscles alters the myosin heavy chain profile permanently. This study provides a rationale to also consider biological solutions to improve nerve regeneration and reinnervation in the surgical repair of lacerated muscles.


Assuntos
Músculo Esquelético/lesões , Músculo Esquelético/inervação , Cadeias Pesadas de Miosina/análise , Acetilcolinesterase/análise , Adenosina Trifosfatases/metabolismo , Animais , Placa Motora/enzimologia , Músculo Esquelético/química , Regeneração Nervosa , Isoformas de Proteínas , Coelhos
3.
Tissue Cell ; 40(3): 219-28, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18242655

RESUMO

It was recently demonstrated that osteogenesis of hESC was more efficient without the initial embryoid body formation step. This study sought to further improve this direct differentiation culture system, by developing an autologous osteogenic-inducing culture supplement extracted from hESC-derived osteogenic cells themselves. A whole cell lysate was prepared from hESC-derived osteogenic cells, simply by exposure to deionized water followed by free-thawing and subsequent filtration. The product was used to coat the surface of cell culture dishes together with gelatin, prior to culture of hESC under osteogenic-inducing conditions. The results showed that the autologous cell lysate extract promoted the aggregation and clustering of cells to form nodule-like structures. Immunohistochemical staining on day 9 demonstrated that these cellular aggregates strongly expressed STRO-1, while on day 14 the nodule-like structures stained positively for both osteocalcin and osteonectin (SPARC). By contrast, the negative control (gelatin coating alone) showed much less prominent cellular aggregation and clustering, and was stained much less intensely for these markers. Additionally, Von Kossa staining on day 14 was also more intense in the presence of the autologous cell lysate extract. Hence, this product can be used to further enhance the osteogenesis of hESC. This would save costs from the use of highly-expensive cytokines, growth factors and matrix components, as well as avoid pathogenic transmission from animal and human products.


Assuntos
Células-Tronco Embrionárias/citologia , Osteoblastos/citologia , Osteogênese , Antígenos de Superfície/metabolismo , Diferenciação Celular , Extratos Celulares , Forma Celular , Células Cultivadas , Humanos , Imuno-Histoquímica , Osteocalcina/metabolismo , Osteonectina/metabolismo
4.
J Orthop Res ; 24(1): 102-11, 2006 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-16419975

RESUMO

The recovery of lacerated skeletal muscles are said to be slow and incomplete. Often the intramuscular (IM-) nerve is concomitantly cut, but never repaired. We questioned whether the IM-nerve should also be reanastamosed before repairing the skeletal muscle. Before answering this, it was necessary to know if the cut IM nerve would have an effect on the recovery of the segment of muscle distal to the level of the laceration. This study investigates the recovery of lacerated muscles after repair, and compares a complete muscle laceration where the main IM-nerve was concomitantly cut and an incomplete muscle laceration where the IM-nerve was preserved intact. The medial gastrocnemius (MG) of the adult male New Zealand White rabbit was used, with the contralateral muscle as a sham control. The laceration was at the proximal quarter of the muscle, distal to the entry point of the nerve branch from the tibial nerve into the muscle belly. Twenty-eight weeks post-repair, the lacerated MG with the IM-nerve intact showed improved muscle wet weight, near normal morphology and contractile properties, and return of muscle fiber type mix and size. The repaired lacerated MG with their IM-nerve concomitantly cut demonstrated loss of muscle wet weight, obvious fibrosis, mononuclear proliferation with fatty infiltration, increase in type-1 fibers and muscle fiber atrophy in the distal portion. We postulate that it might be important to repair the intramuscular nerve branch by microanastomosis when repairing a vital skeletal muscle that is lacerated.


Assuntos
Músculo Esquelético/lesões , Músculo Esquelético/inervação , Cicatrização , Animais , Estimulação Elétrica , Masculino , Contração Muscular/fisiologia , Fibras Musculares Esqueléticas/patologia , Músculo Esquelético/patologia , Coelhos , Nervo Tibial/lesões
5.
Muscle Nerve ; 33(3): 377-83, 2006 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-16320308

RESUMO

The repair of lacerated muscle often results in suboptimal recovery. An important cause of poor outcome is denervation of the distal segment. The rabbit medial gastrocnemius muscle laceration model was used to assess whether intramuscular nerve repair resulted in better recovery. Lacerated rabbit muscles were divided into three groups: group A had no muscle repair; group B underwent muscle repair; and group C had muscle repair with intramuscular nerve repair. At 7 months, groups A and B showed significantly greater muscle atrophy, replacement of muscle fiber with scar and adipose tissue, and change of muscle fiber type from a fast-twitch to a slow-twitch pattern compared to group C. A clinical case study subsequently demonstrated feasibility of intramuscular nerve repair; reinnervation of the distal belly led to rapid functional recovery. In conclusion, primary intramuscular nerve repair results in better functional outcomes following repair of lacerated muscles.


Assuntos
Lacerações/patologia , Lacerações/cirurgia , Músculo Esquelético/lesões , Músculo Esquelético/inervação , Procedimentos Neurocirúrgicos , Adenosina Trifosfatases/metabolismo , Tecido Adiposo/patologia , Adulto , Animais , Peso Corporal/fisiologia , Cicatriz/patologia , Eletromiografia , Fibrose , Humanos , Masculino , Denervação Muscular , Fibras Musculares Esqueléticas/patologia , Músculo Esquelético/patologia , Atrofia Muscular/patologia , Tamanho do Órgão/fisiologia , Coelhos , Resultado do Tratamento
6.
J Orthop Res ; 22(6): 1325-30, 2004 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-15475216

RESUMO

Bipennate muscles may be split along their distal aponeurosis, dividing each into two compartments. These sub-muscle units may be used in tendon transfers. This paper presents the contractile properties of the two sub-units of the flexor carpi ulnaris in a macaca fascicularis, after it was split by up to 80% of its length. The sub-muscle units were electrically stimulated and found to have independent isometric contraction, with minimal contraction recorded from the non-stimulated sub-unit. Also, the sum of the forces measured from each unit when stimulated individually, was found to be greater than the force of the whole muscle, given the same isometric conditions. The distal aponeurosis which is common allows force transmission between the compartments. Splitting the muscle along this distal aponeurosis alters this function and the force capacity of the muscle, providing a new potential for using the sub-units as grafts for tendon transfers.


Assuntos
Contração Isométrica/fisiologia , Músculo Esquelético/inervação , Músculo Esquelético/fisiologia , Tendões/fisiologia , Animais , Estimulação Elétrica , Macaca fascicularis , Músculo Esquelético/cirurgia , Procedimentos de Cirurgia Plástica , Tendões/cirurgia , Nervo Ulnar/fisiologia
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