Chondrogenesis of cocultures of mesenchymal stem cells and articular chondrocytes in poly(l-lysine)-loaded hydrogels

Kim, Yu Seon; Chien, Athena J.; Guo, Jason L; Smith, Brandon T.; Watson, Emma; Pearce, Hannah A.; Koons, Gerry L.; Navara, Adam M.; Lam, Johnny; Scott, David W.; Grande‐Allen, K. Jane

doi:10.1016/j.jconrel.2020.09.048

Cited by 14 publications

(37 citation statements)

References 34 publications

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“…Following hydrogel formation, the swelling and degradation behavior were assessed as previously described [ 1 , 4 , 9 , 11 , 14 , 27 , 29 , 30 ]. After crosslinking for 24 h at 37°C to form the hydrogels, the hydrogels were transferred and weighed in a pre-weighed 20 ml scintillation vial to obtain the formation weight ( W f ).…”

Section: Methodsmentioning

confidence: 99%

Evaluating the physicochemical effects of conjugating peptides into thermogelling hydrogels for regenerative biomaterials applications

Pearce

Jiang

Swain

et al. 2021

Regenerative Biomaterials

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Thermogelling hydrogels such as poly(N-isopropyl acrylamide) (P(NiPAAm)) provide tunable constructs leveraged in many regenerative biomaterial applications. Recently, our lab developed the crosslinker poly(glycolic acid)-poly(ethylene glycol)-poly(glycolic acid)-di(but-2-yne-1,4-dithiol) (PdBT), which crosslinks P(NiPAAm-co-glycidyl methacrylate) via thiol-epoxy reaction and can be functionalized with azide-terminated peptides via alkyne-azide click chemistry. This study’s aim was to evaluate the impact of peptides on the physicochemical properties of the hydrogels. The physicochemical properties of the hydrogels including the lower critical solution temperature, crosslinking times, swelling, degradation, peptide release, and cytocompatibility were evaluated. The gels bearing peptides increased equilibrium swelling indicating hydrophilicity of the hydrogel components. Comparable sol fractions were found for all groups, indicating that inclusion of peptides does not impact crosslinking. Moreover, the inclusion of a matrix metalloproteinase (MMP)-sensitive peptide allowed elucidation of whether release of peptides from the network was driven by hydrolysis or enzymatic cleavage. The hydrophilicity of the network determined by the swelling behavior was demonstrated to be the most important factor in dictating hydrogel behavior over time. This study demonstrates the importance of characterizing the impact of additives on the physicochemical properties of hydrogels. These characteristics are key in determining design considerations for future in vitro and in vivo studies for tissue regeneration.

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Section: Methodsmentioning

confidence: 99%

Evaluating the physicochemical effects of conjugating peptides into thermogelling hydrogels for regenerative biomaterials applications

Pearce

Jiang

Swain

et al. 2021

Regenerative Biomaterials

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“…PLL MW and concentration of 225 kDa and 50 μg/ml were used for all PLL‐loaded study groups. The values were selected in a previous study conducted in our laboratory (Kim et al, 2020). All study groups were processed for same sets of experiments to determine the behaviors of encapsulated cells (proliferation and sGAG secretion) as well the mechanical properties of hydrogels injected within the cartilage explants.…”

Section: Methodsmentioning

confidence: 99%

“…DNA and sGAG content were quantified at each time point (Weeks 1 and 3) following a previously established protocol (Kim et al, 2020).…”

Section: Biochemical Analysesmentioning

confidence: 99%

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Evaluation of tissue integration of injectable, cell‐laden hydrogels of cocultures of mesenchymal stem cells and articular chondrocytes with an ex vivo cartilage explant model

Kim

Mehta

Guo

et al. 2021

Biotech & Bioengineering

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This study investigated the chondrogenic activity of encapsulated mesenchymal stem cells (MSCs) and articular chondrocytes (ACs) and its impact on the mechanical properties of injectable poly(N-isopropylacrylamide)-based dual-network hydrogels loaded with poly(L-lysine) (PLL). To this effect, an ex vivo study model was employed to assess the behavior of the injected hydrogels-specifically, their surface stiffness and integration strength with the surrounding cartilage. The highest chondrogenic activity was observed from AC-encapsulated hydrogels, while the effect of PLL on MSC chondrogenesis was not apparent from biochemical analyses. Mechanical testing showed that there were no significant differences in either surface stiffness or integration strength among the different study groups. Altogether, the results suggest that the ex vivo model can allow further understanding of the relationship between biochemical changes within the hydrogel and their impact on the hydrogel's mechanical properties.

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“…During the past decade, tissue engineering has shown promising clinical application prospects in cartilage repair. Poly L-lysine (PLL), a polypeptide with cationic lysine groups as its functional group, has been shown to enhance chondrogenesis in a dose and molecular weight dependent manner in vitro (40). However, the underlying mechanisms of circRNAs in these signaling pathways in KOA remain unclear, which need further study.…”

Section: Qrt-pcr Validation Of Hsa_circ_0005265 In Both Synovium and Ipfpmentioning

confidence: 99%

RNA sequencing reveals the circular RNA expression profiles of the infrapatellar fat pad/synovium unit

Jiang¹,

Lu²,

Chen³

et al. 2021

Ann Transl Med

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Background: The infrapatellar fat pad (IPFP) and synovium are reported to act as a functional unit, with emerging roles in the pathophysiology of knee osteoarthritis (KOA). Circular RNAs (circRNAs) are involved in the pathogenesis of KOA, especially in cartilage homeostasis regulation. Nevertheless, the regulatory mechanisms of circRNAs in the KOA IPFP/synovium unit remain to be elucidated. Therefore, the current study aimed to investigate alterations in the expression of circRNAs and predict their functions in the KOA IPFP/synovium unit using bioinformatics analysis.Methods: In brief, 6 synovium and IPFP specimens were collected, in which 3 from patients with KOA and 3 from controls. Then, circRNA sequencing was conducted on 2 KOA synovium and IPFP specimens as well as 1 control to investigate the expression profiles of circRNAs. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Reactome signaling pathway analyses were employed to predict the functions of the differentially expressed circRNAs. Based on the miRNA sponge theory, we constructed a circRNA-miRNA network to predict the molecular regulatory mechanism of these circRNAs. Venn analysis was performed to confirm the circRNAs and miRNAs expressed in both synovium and IPFP. Realtime quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was also used to validate the expression levels of circRNAs that were co-expressed in both synovium and IPFP.Results: A total of 65 and 72 circRNAs were differentially expressed in KOA synovium and IPFP, respectively (fold change ≥2, P<0.05). After obtaining the parental genes of differentially expressed circRNAs, the top 10 enrichment GO entries, KEGG pathways, and Reactome pathways were annotated. Furthermore, hsa_circ_0005265 was found to be down-regulated in both synovium and IPFP, which was validated by qRT-PCR. The circRNA-miRNA network was created to annotate the probable regulatory mechanisms of the differentially expressed circRNAs, which consequently confirmed 2 target miRNAs (hsa-miR-6769b-5p and hsa-miR-1249-5p) associated with hsa_circ_0005265 in both synovium and IPFP.Conclusions: Our outcomes bring us closer to understanding the potential mechanism of the IPFP/ synovium unit in the progression of KOA and finding new molecular targets for KOA therapy.

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Chondrogenesis of cocultures of mesenchymal stem cells and articular chondrocytes in poly(l-lysine)-loaded hydrogels

Cited by 14 publications

References 34 publications

Evaluating the physicochemical effects of conjugating peptides into thermogelling hydrogels for regenerative biomaterials applications

Evaluating the physicochemical effects of conjugating peptides into thermogelling hydrogels for regenerative biomaterials applications

Evaluation of tissue integration of injectable, cell‐laden hydrogels of cocultures of mesenchymal stem cells and articular chondrocytes with an ex vivo cartilage explant model

RNA sequencing reveals the circular RNA expression profiles of the infrapatellar fat pad/synovium unit

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