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Journal of Molecular Histology

, Volume 49, Issue 4, pp 339–345 | Cite as

Three-dimensional reconstructed eccrine sweat glands with vascularization and cholinergic and adrenergic innervation

  • Mingjun Zhang
  • Haihong Li
  • Liyun Chen
  • Shuhua Fang
  • Sitian Xie
  • Changmin Lin
Original Paper
  • 127 Downloads

Abstract

Functional integrity of the regenerated tissues requires not only structural integrity but also vascularization and innervation. We previously demonstrated that the three-dimensional (3D) reconstructed eccrine sweat glands had similar structures as those of the native ones did, but whether the 3D reconstructed glands possessing vascularization and innervation was still unknown. In the study, Matrigel-embedded eccrine sweat gland cells were implanted under the inguinal skin. Ten weeks post-implantation, the vascularization, and innervation in the 10-week reconstructed eccrine sweat glands and native human eccrine sweat glands were detected by immunofluorescence staining. The results showed that the fluorescent signals of general neuronal marker protein gene product 9.5, adrenergic nerve fiber marker tyrosine hydroxylase, and cholinergic nerve fiber markers acetylcholinesterase and vasoactive intestinal peptide embraced the 3D reconstructed glands in circular patterns, as the signals appeared in native eccrine sweat glands. There were many CD31- and von Willebrand factor-positive vessels growing into the plugs. We demonstrated that the 3D reconstructed eccrine sweat glands were nourished by blood vessels, and we for the first time demonstrated that the engineering sweat glands were innervated by both cholinergic and adrenergic fibers. In conclusion, the 3D reconstructed eccrine sweat glands may have functions as the native ones do.

Keywords

Eccrine sweat gland Three-dimensional reconstruction Matrigel Innervation Vascularization 

Abbreviations

AChE

Acetylcholinesterase

DAPI

4′,6-diamidino-2-phenylindole

FITC

Fluorescein isothiocyanate

HE

Hematoxylin-eosin

KSFM

Defined keratinocyte serum-free medium

HRP

Horseradish peroxidase

PBS

Phosphate-buffered saline

PGP 9.5

Protein gene product 9.5

SUMC

Shantou University Medical College

TH

Tyrosine hydroxylase

TSA

Tyramide signal amplification

VIP

Vasoactive intestinal peptide

vWF

Von Willebrand factor

Notes

Acknowledgements

The manuscript was supported in part by the National Natural Science Foundation of China (81772102, 81471882).

Compliance with ethical standards

Conflict of interest

We declare we have no competing financial, personal or other relationships with other people or organizations.

References

  1. Cowen T, Thrasivoulou C, Shaw SA, Abdel-Rahman TA (1996) Transplanted sweat glands from mature and aged donors determine cholinergic phenotype and altered density of host sympathetic nerves. J Auton Nerv Syst 60:215–224CrossRefPubMedGoogle Scholar
  2. Danner S, Kremer M, Petschnik AE, Nagel S, Zhang Z, Hopfner U, Reckhenrich AK, Weber C, Schenck TL, Becker T, Kruse C, Machens HG, Egana JT (2012) The use of human sweat gland-derived stem cells for enhancing vascularization during dermal regeneration. J Invest Dermatol 132:1707–1716CrossRefPubMedGoogle Scholar
  3. Distler JH, Hirth A, Kurowska-Stolarska M, Gay RE, Gay S, Distler O (2003) Angiogenic and angiostatic factors in the molecular control of angiogenesis. Q J Nucl Med 47:149–161PubMedGoogle Scholar
  4. Fu X, Li X, Cheng B, Chen W, Sheng Z (2005a) Engineered growth factors and cutaneous wound healing: success and possible questions in the past 10 years. Wound Repair Regen 13:122–130CrossRefPubMedGoogle Scholar
  5. Fu XB, Sun TZ, Li XK, Sheng ZY (2005b) Morphological and distribution characteristics of sweat glands in hypertrophic scar and their possible effects on sweat gland regeneration. Chin Med J (Engl) 118:186–191Google Scholar
  6. Gao LP, Du MJ, Lv JJ, Schmull S, Huang RT, Li J (2017) Use of human aortic extracellular matrix as a scaffold for construction of a patient-specific tissue engineered vascular patch. Biomed Mater 12:065006CrossRefPubMedGoogle Scholar
  7. Huging M, Biedermann T, Sobrio M, Meyer S, Bottcher-Haberzeth S, Manuel E, Horst M, Hynes S, Reichmann E, Schiestl C, Hartmann-Fritsch F (2017) The effect of wound dressings on a bio-engineered human dermo-epidermal skin substitute in a rat model. J Burn Care Res 38:354–364CrossRefGoogle Scholar
  8. Karsy M, Burnett B, Di Ieva A, Cusimano MD, Jensen RL (2018) Microvascularization of Grade I meningiomas: effect on tumor volume, blood loss, and patient outcome. J Neurosurg 128(3):657–666.  https://doi.org/10.3171/2016.10.JNS161825 CrossRefPubMedGoogle Scholar
  9. Klar AS, Michalak K, Böttcher-Haberzeth S, Reichmann E, Meuli M, Biedermann T (2018) The expression pattern of keratin 24 in tissue-engineered dermo-epidermal human skin substitutes in an in vivo model. Pediatr Surg Int 34(2):237–244.  https://doi.org/10.1007/s00383-017-4198-9 CrossRefPubMedGoogle Scholar
  10. Landis SC, Fredieu JR (1986) Coexistence of calcitonin gene-related peptide and vasoactive intestinal peptide in cholinergic sympathetic innervation of rat sweat glands. Brain Res 377:177–181CrossRefPubMedGoogle Scholar
  11. Li H, Chen L, Zeng S, Li X, Zhang X, Lin C, Zhang M, Xie S, He Y, Shu S, Yang L, Tang S, Fu X (2015) Matrigel basement membrane matrix induces eccrine sweat gland cells to reconstitute sweat gland-like structures in nude mice. Exp Cell Res 332:67–77CrossRefPubMedGoogle Scholar
  12. Li H, Li X, Zhang B, Zhang M, Chen W, Tang S, Fu X (2016a) Changes in keratins and alpha-smooth muscle actin during three-dimensional reconstitution of eccrine sweat glands. Cell Tissue Res 365:113–122CrossRefPubMedGoogle Scholar
  13. Li H, Zhang M, Chen L, Li X, Zhang B (2016b) Human eccrine sweat gland cells reconstitute polarized spheroids when subcutaneously implanted with Matrigel in nude mice. J Mol Histol 47:485–490CrossRefPubMedGoogle Scholar
  14. Li H, Chen L, Zhang M, Zhang B (2017a) Foxa1 gene and protein in developing rat eccrine sweat glands. J Mol Histol 48:1–7CrossRefPubMedGoogle Scholar
  15. Li H, Zhang M, Chen L, Zhang B, Zhang C (2017b) Expression of S100A2 and S100P in human eccrine sweat glands and their application in differentiating secretory coil-like from duct-like structures in the 3D reconstituted eccrine sweat spheroids. J Mol Histol 48:219–223CrossRefPubMedGoogle Scholar
  16. Li X, Li H, Zhang M, Chen L, Zhang B (2017c) Cell proliferation and differentiation during the three dimensional reconstitution of eccrine sweat glands. J Mol Histol 48:113–120CrossRefPubMedGoogle Scholar
  17. Lu CP, Polak L, Keyes BE, Fuchs E (2016) Spatiotemporal antagonism in mesenchymal-epithelial signaling in sweat versus hair fate decision. Science 354(6319):aah6102.  https://doi.org/10.1126/science.aah6102 CrossRefPubMedPubMedCentralGoogle Scholar
  18. Rittie L, Sachs DL, Orringer JS, Voorhees JJ, Fisher GJ (2013) Eccrine sweat glands are major contributors to reepithelialization of human wounds. Am J Pathol 182:163–171CrossRefPubMedPubMedCentralGoogle Scholar
  19. Saga K (2002) Structure and function of human sweat glands studied with histochemistry and cytochemistry. Prog Histochem Cytochem 37:323–386CrossRefPubMedGoogle Scholar
  20. Schotzinger RJ, Landis SC (1990) Acquisition of cholinergic and peptidergic properties by sympathetic innervation of rat sweat glands requires interaction with normal target. Neuron 5:91–100CrossRefPubMedGoogle Scholar
  21. Schotzinger R, Yin X, Landis S (1994) Target determination of neurotransmitter phenotype in sympathetic neurons. J Neurobiol 25:620–639CrossRefPubMedGoogle Scholar
  22. Sheng Z, Fu X, Cai S, Lei Y, Sun T, Bai X, Chen M (2009) Regeneration of functional sweat gland-like structures by transplanted differentiated bone marrow mesenchymal stem cells. Wound Repair Regen 17:427–435CrossRefPubMedGoogle Scholar
  23. Stanke M, Duong CV, Pape M, Geissen M, Burbach G, Deller T, Gascan H, Otto C, Parlato R, Schutz G, Rohrer H (2006) Target-dependent specification of the neurotransmitter phenotype: cholinergic differentiation of sympathetic neurons is mediated in vivo by gp 130 signaling. Development 133:141–150CrossRefPubMedGoogle Scholar
  24. Stevens LM, Landis SC (1987) Development and properties of the secretory response in rat sweat glands: relationship to the induction of cholinergic function in sweat gland innervation. Dev Biol 123:179–190CrossRefPubMedGoogle Scholar
  25. Stevens LM, Landis SC (1988) Developmental interactions between sweat glands and the sympathetic neurons which innervate them: effects of delayed innervation on neurotransmitter plasticity and gland maturation. Dev Biol 130:703–720CrossRefPubMedGoogle Scholar
  26. Uno H, Hokfelt T (1975) Catecholamine-containing nerve terminals of the eccrine sweat glands of macaques. Cell Tissue Res 158:1–13CrossRefPubMedGoogle Scholar
  27. Wang N, Gibbons CH, Freeman R (2011) Novel immunohistochemical techniques using discrete signal amplification systems for human cutaneous peripheral nerve fiber imaging. J Histochem Cytochem 59:382–390CrossRefPubMedPubMedCentralGoogle Scholar
  28. Watt SM, Pleat JM (2018) Stem cells, niches and scaffolds: applications to burns and wound care. Adv Drug Deliv Rev 123:82–106.  https://doi.org/10.1016/j.addr.2017.10.012 CrossRefPubMedGoogle Scholar
  29. Zhang C, Chen Y, Fu X (2015) Sweat gland regeneration after burn injury: is stem cell therapy a new hope? Cytotherapy 17:526–535CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media B.V., part of Springer Nature 2018

Authors and Affiliations

  • Mingjun Zhang
    • 1
  • Haihong Li
    • 1
  • Liyun Chen
    • 1
  • Shuhua Fang
    • 1
  • Sitian Xie
    • 1
  • Changmin Lin
    • 2
  1. 1.Department of Burn and Plastic Surgery, The Second Affiliated HospitalShantou University Medical CollegeShantouChina
  2. 2.Department of Histology and EmbryologyShantou University Medical CollegeShantouChina

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