Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold
Due to a relative paucity of studies on human lymphatic assembly in vitro and subsequent in vivo transplantation, capillary formation and survival of primary human lymphatic (hLEC) and blood endothelial cells (hBEC) ± primary human vascular smooth muscle cells (hvSMC) were evaluated and compared in...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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SAGE Publishing
2022-12-01
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| Series: | Journal of Tissue Engineering |
| Online Access: | https://doi.org/10.1177/20417314221140979 |
| _version_ | 1797976742272434176 |
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| author | Anne M Kong Shiang Y Lim Jason A Palmer Amanda Rixon Yi-Wen Gerrand Kiryu K Yap Wayne A Morrison Geraldine M Mitchell |
| author_facet | Anne M Kong Shiang Y Lim Jason A Palmer Amanda Rixon Yi-Wen Gerrand Kiryu K Yap Wayne A Morrison Geraldine M Mitchell |
| author_sort | Anne M Kong |
| collection | DOAJ |
| description | Due to a relative paucity of studies on human lymphatic assembly in vitro and subsequent in vivo transplantation, capillary formation and survival of primary human lymphatic (hLEC) and blood endothelial cells (hBEC) ± primary human vascular smooth muscle cells (hvSMC) were evaluated and compared in vitro and in vivo. hLEC ± hvSMC or hBEC ± hvSMC were seeded in a 3D porous scaffold in vitro, and capillary percent vascular volume (PVV) and vascular density (VD)/mm 2 assessed. Scaffolds were also transplanted into a sub-cutaneous rat wound with morphology/morphometry assessment. Initially hBEC formed a larger vessel network in vitro than hLEC, with interconnected capillaries evident at 2 days. Interconnected lymphatic capillaries were slower (3 days) to assemble. hLEC capillaries demonstrated a significant overall increase in PVV ( p = 0.0083) and VD ( p = 0.0039) in vitro when co-cultured with hvSMC. A similar increase did not occur for hBEC + hvSMC in vitro, but hBEC + hvSMC in vivo significantly increased PVV ( p = 0.0035) and VD ( p = 0.0087). Morphology/morphometry established that hLEC vessels maintained distinct cell markers, and demonstrated significantly increased individual vessel and network size, and longer survival than hBEC capillaries in vivo, and established inosculation with rat lymphatics, with evidence of lymphatic function. The porous polyurethane scaffold provided advantages to capillary network formation due to its large (300–600 μm diameter) interconnected pores, and sufficient stability to ensure successful surgical transplantation in vivo. Given their successful survival and function in vivo within the porous scaffold, in vitro assembled hLEC networks using this method are potentially applicable to clinical scenarios requiring replacement of dysfunctional or absent lymphatic networks. |
| first_indexed | 2024-04-11T04:55:48Z |
| format | Article |
| id | doaj.art-b602b3a137584812bf6eb329c7df97b7 |
| institution | Directory Open Access Journal |
| issn | 2041-7314 |
| language | English |
| last_indexed | 2024-04-11T04:55:48Z |
| publishDate | 2022-12-01 |
| publisher | SAGE Publishing |
| record_format | Article |
| series | Journal of Tissue Engineering |
| spelling | doaj.art-b602b3a137584812bf6eb329c7df97b72022-12-26T15:33:48ZengSAGE PublishingJournal of Tissue Engineering2041-73142022-12-011310.1177/20417314221140979Engineering transplantable human lymphatic and blood capillary networks in a porous scaffoldAnne M Kong0Shiang Y Lim1Jason A Palmer2Amanda Rixon3Yi-Wen Gerrand4Kiryu K Yap5Wayne A Morrison6Geraldine M Mitchell7O’Brien Institute Department of St Vincent’s Institute of Medical Research, Fitzroy, VIC, AustraliaNational Heart Research Institute Singapore, National Heart Centre SingaporeCentre for Eye Research Australia, East Melbourne, VIC, AustraliaExperimental Medical and Surgical Unit, St Vincent’s Hospital Melbourne, Fitzroy, VIC, AustraliaO’Brien Institute Department of St Vincent’s Institute of Medical Research, Fitzroy, VIC, AustraliaDepartment of Surgery at St Vincent’s Hospital Melbourne, University of Melbourne, Fitzroy, VIC, AustraliaDepartment of Plastic and Reconstructive Surgery, St Vincent’s Hospital Melbourne, Fitzroy, VIC, AustraliaFaculty of Health Sciences, Australian Catholic University, East Melbourne VIC, AustraliaDue to a relative paucity of studies on human lymphatic assembly in vitro and subsequent in vivo transplantation, capillary formation and survival of primary human lymphatic (hLEC) and blood endothelial cells (hBEC) ± primary human vascular smooth muscle cells (hvSMC) were evaluated and compared in vitro and in vivo. hLEC ± hvSMC or hBEC ± hvSMC were seeded in a 3D porous scaffold in vitro, and capillary percent vascular volume (PVV) and vascular density (VD)/mm 2 assessed. Scaffolds were also transplanted into a sub-cutaneous rat wound with morphology/morphometry assessment. Initially hBEC formed a larger vessel network in vitro than hLEC, with interconnected capillaries evident at 2 days. Interconnected lymphatic capillaries were slower (3 days) to assemble. hLEC capillaries demonstrated a significant overall increase in PVV ( p = 0.0083) and VD ( p = 0.0039) in vitro when co-cultured with hvSMC. A similar increase did not occur for hBEC + hvSMC in vitro, but hBEC + hvSMC in vivo significantly increased PVV ( p = 0.0035) and VD ( p = 0.0087). Morphology/morphometry established that hLEC vessels maintained distinct cell markers, and demonstrated significantly increased individual vessel and network size, and longer survival than hBEC capillaries in vivo, and established inosculation with rat lymphatics, with evidence of lymphatic function. The porous polyurethane scaffold provided advantages to capillary network formation due to its large (300–600 μm diameter) interconnected pores, and sufficient stability to ensure successful surgical transplantation in vivo. Given their successful survival and function in vivo within the porous scaffold, in vitro assembled hLEC networks using this method are potentially applicable to clinical scenarios requiring replacement of dysfunctional or absent lymphatic networks.https://doi.org/10.1177/20417314221140979 |
| spellingShingle | Anne M Kong Shiang Y Lim Jason A Palmer Amanda Rixon Yi-Wen Gerrand Kiryu K Yap Wayne A Morrison Geraldine M Mitchell Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold Journal of Tissue Engineering |
| title | Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold |
| title_full | Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold |
| title_fullStr | Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold |
| title_full_unstemmed | Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold |
| title_short | Engineering transplantable human lymphatic and blood capillary networks in a porous scaffold |
| title_sort | engineering transplantable human lymphatic and blood capillary networks in a porous scaffold |
| url | https://doi.org/10.1177/20417314221140979 |
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