N-cadherin is Key to Expression of the Nucleus Pulposus Cell Phenotype under Selective Substrate Culture Conditions.
Nucleus pulposus (NP) cells of the intervertebral disc are essential for synthesizing extracellular matrix that contributes to disc health and mechanical function. NP cells have a unique morphology and molecular expression pattern derived from their notochordal origin, and reside in N-cadherin (CDH2) positive cell clusters in vivo. With disc degeneration, NP cells undergo morphologic and phenotypic changes including loss of CDH2 expression and ability to form cell clusters. Here, we investigate the role of CDH2 positive cell clusters in preserving healthy, biosynthetically active NP cells. Using a laminin-functionalized hydrogel system designed to mimic features of the native NP microenvironment, we demonstrate NP cell phenotype and morphology is preserved only when NP cells form CDH2 positive cell clusters. Knockdown (CRISPRi) or blocking CDH2 expression in vitro and in vivo results in loss of a healthy NP cell. Findings also reveal that degenerate human NP cells that are CDH2 negative can be promoted to re-express CDH2 and healthy, juvenile NP matrix synthesis patterns by promoting cell clustering for controlled microenvironment conditions. This work also identifies CDH2 interactions with β-catenin-regulated signaling as one mechanism by which CDH2-mediated cell interactions can control NP cell phenotype and biosynthesis towards maintenance of healthy intervertebral disc tissues.
Department of Biomedical Engineering, Duke University, Durham NC 27713, USA.|Department of Biomedical Engineering, Washington University in St. Louis, St. Louis Missouri 63130, USA.
Priscilla Y
PY
Hwang
Department of Biomedical Engineering, Duke University, Durham NC 27713, USA.|Department of Biomedical Engineering, Washington University in St. Louis, St. Louis Missouri 63130, USA.
Liufang
L
Jing
Department of Orthopaedic Surgery, Duke University Medical Center, Durham NC 27713, USA.
Jun
J
Chen
Departments of Orthopaedics &Traumatology, The University of Hong Kong, Hong Kong SAR, China.
Foon-Lian
FL
Lim
Department of Orthopaedic Surgery, Washington University in St. Louis, St. Louis Missouri 63110, USA.
Ruhang
R
Tang
Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia PA, USA.
Hyowon
H
Choi
Departments of Orthopaedics &Traumatology, The University of Hong Kong, Hong Kong SAR, China.
Kenneth M
KM
Cheung
Department of Orthopaedic Surgery, Thomas Jefferson University, Philadelphia PA, USA.
Makarand V
MV
Risbud
Department of Biomedical Engineering, Duke University, Durham NC 27713, USA.
Charles A
CA
Gersbach
Department of Biomedical Engineering, Duke University, Durham NC 27713, USA.|Department of Biomedical Engineering, Washington University in St. Louis, St. Louis Missouri 63130, USA.|Department of Orthopaedic Surgery, Washington University in St. Louis, St. Louis Missouri 63110, USA.
Farshid
F
Guilak
Departments of Orthopaedics &Traumatology, The University of Hong Kong, Hong Kong SAR, China.
Victor Y
VY
Leung
Department of Biomedical Engineering, Duke University, Durham NC 27713, USA.|Department of Biomedical Engineering, Washington University in St. Louis, St. Louis Missouri 63130, USA.|Department of Orthopaedic Surgery, Washington University in St. Louis, St. Louis Missouri 63110, USA.
Lori A
LA
Setton
D000293:Adolescent; D000328:Adult; D000368:Aged; D000818:Animals; D015703:Antigens, CD / Q000235:genetics* / Q000378:metabolism*; D015820:Cadherins / Q000235:genetics* / Q000378:metabolism*; D002450:Cell Communication; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D002648:Child; D055785:Gene Knockdown Techniques; D006801:Humans; D008875:Middle Aged; D000070614:Nucleus Pulposus / Q000166:cytology* / Q000378:metabolism; D010641:Phenotype; D015398:Signal Transduction; D013552:Swine; D055815:Young Adult; D051176:beta Catenin / Q000378:metabolism
D015703:Antigens, CD; C050361:CDH2 protein, human; D015820:Cadherins; D051176:beta Catenin