Research Article Open Access Novel Physiotherapies Hadjileontis and Kontakis, J Nov Physiother 2013, S2 http://dx.doi.org/10.4172/2165-7025.S2-007 J Nov Physiother Orthopedics and Sports Physical Therapy ISSN:2165-7025 JNP, an open access journal Evidence of Neuronal Differentiation of Tendon Stromal Cells in Patients with Biceps Branchi Muscle Pain: A Histological and Immunohistochemical Study of 12 Patients Constantine Hadjileontis 1 * and George Kontakis 2 1 Pathology Department of Serres General Hospital, Serres, Greece 2 Department of Orthopaedic Surgery, Heraklion University Hospital, Crete, Greece Keywords: Long biceps tendon; Pain; Histology- immunohistochemistry Introduction Te long head of the biceps branchii is a gliding tendon, which allows the humeral head to move on the fxed tendon during motion at the shoulder joint. In order to investigate the actual cause of pain afer shoulder fractures and injuries, we histologically examined the lesions present at the tendon, afer sampling during surgery reconstruction. Tis would enable the clarifcation of the mechanisms involved in pain generation, and the detection of targeted pharmaceutical schemes, to achieve long term post-operative pain restriction [1,2]. Until now very few studies concerning the histological lesions of biceps branchii tendons are released [3-5]. Results from these studies show that afected tendons exhibit marked histological changes, compared with normal tendons of both younger and aged individuals [3]. Physiologically tendons show close and parallel arrangement of collagen fbers with slight waviness, that provide a deep pink-red staining result afer treated with hematoxylin and eosin (H&E). Tese fbers include tenocytes with fattened and spindle shaped nuclei, ofen arranged in rows. Tese cells represent modifed, inactive fbroblasts. No neural (sensor) cells are normally present within the tendon-fber matrix (tendon tissue proper), while blood vessels and nerves are supplied from the surrounding tissues (paratendinous), branching up to a point into the periphery of tendon tissue proper, parallel to its collagen fbers [4]. What’s new in this study is that all morphologic measurements were performed quantitatively and tendons were dissected in three parts (proximal, middle and distal) that were separately studied for each lesion. Furthermore, we tried to immunohistochemically detect cells with neural features and function within tendon tissue proper, that would explain pain manifestation. We also examined possible mechanisms that might stimulate and/or sustain the appearance of these cells, such as infammation and neoangiogenesis. In order to achieve these goals we applied well-documented histological criteria for each lesion that are thoroughly substantiated in literature. Materials and Methods Tendons from all cases (n=12) were dissected in three parts and were oriented as proximal, middle and distal edge, according to their anatomical relation with the afected joint. Biopsies were then fxed in formalin bufer 4% for 24 h in order to proceed to the histological and immunohistochemical evaluation. Te lesions studied included tendon cellularity and vascularity, the presence and density of infammation (both histologically and immunohistochemically by using common leukocyte antigen), the degree and type of tendon degeneration, as well as the expression of S-100 protein and neuropeptide Y in the afected tissues. For the angiogenic evaluation we applied Burkhardt’s histological criteria [1] for neo-angiogenesis. Te immunohistochemical marker used for staining endothelial cells was CD34. Morphological evaluation was performed afer applying the routine stain of H&E, whereas for immunohistochemistry we used the Vision Biosystem Bond max staining machine, prior to applying the immunohistochemical markers (Table 1). Tis staining machine includes steps to dewax, pretreat and stain slides, using a primary antibody that attaches to the tissue antigens. A polymer- Abstract The tendon of the long head of biceps branchii muscles is a major source of pain in the shoulder region. Our objective was to detect histologically and characterize factors that may contribute to biceps tendon pain generation, and subsequently provide a possible explanation of pain origin. This would permit the investigation of targeted pharmaceutical treatments for long term post-operative restriction of pain. Bioptic samples were obtained from 11 patients operated for shoulder fractures and were harvested during shoulder reconstruction operations. As a control group we used biceps tendons from 10 forensic cases with no history of shoulder diseases. Histologic investigation included quantitative measurement of tendon degeneration, cellularity, neoangiogenesis, infammation and metaplasia, as well as immunohistochemical detection of cells with neural differentiation within the tendon tissue proper (S-100 protein and neuropeptide Y). Studied lesions were signifcantly (Mann-Whitney and Wilcox on test) greater in the group of patients compared to the ones found in the control group (p<0,001), while these lesions were statistically proved (Pearson test) to relate with each other, resulting in the neural differentiation of tendon stromal cells, causing thus pain. *Corresponding author: Constantine Hadjileontis, Pathology Department of Serres General Hospital, Crete, Greece, E-mail: conhad@hospser.gr Received February 12, 2013; Accepted April 08, 2013; Published April 10, 2013 Citation: Hadjileontis C, Kontakis G (2013) Evidence of Neuronal Differentiation of Tendon Stromal Cells in Patients with Biceps Branchi Muscle Pain: A Histological and Immunohistochemical Study of 12 Patients. J Nov Physiother S2: 007. doi:10.4172/2165-7025.S2-007 Copyright: © 2013 Hadjileontis C, et al. 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