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Applicability of a Newly Developed Bioassay for Determining Bioactivity of Anti-Inflammatory Compounds in Release Studies − Celecoxib and Triamcinolone Acetonide Released from Novel PLGA-Based Microspheres

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Abstract

Purpose

: To develop a bio-assay for measuring long-term bioactivity of released anti-inflammatory compounds and to test the bioactivity of celecoxib (CXB) and triamcinolone acetonide (TA) released from a new PLGA-based microsphere platform.

Methods

: Human osteoarthritic chondrocytes were plated according to standardized procedures after batch-wise harvest and cultured for 3 days to prevent cell confluency and changes in cell behaviour. Prostaglandin E2 (PGE2) production stimulated by TNFα was used as a parameter of inflammation. A novel microsphere platform based on PTE-functionalised PLGA was used to incorporate CXB and TA. Loaded microspheres were added to transwells overlying the cells, with transfer of the wells to new cell cultures every 3 days. Inhibition of PGE2 production was determined over a period of 21 days.

Results

: PLGA(75:25)-PTE microspheres were prepared and loaded with CXB and TA at 86 and 97% loading efficiency, respectively. In the bioactivity assay, PGE2 levels induced by TNFα were reduced to an average of 30% using microspheres loaded with 0.1 nmol CXB per transwell; with microspheres loaded with 0.1 nmol TA, PGE2 production was initially reduced to 3% and gradually recovered to 30% reduction. At 1 nmol loading, PGE2 was inhibited to 0–7% for CXB-loaded microspheres, and 0–28% for TA-loaded microspheres.

Conclusions

: We present a novel sustained release bioactivity assay which provides an essential link between in vitro buffer-based release kinetics and in vivo application. Novel PLGA-based microspheres loaded with TA and CXB showed efficient anti-inflammatory effects over time.

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Abbreviations

CXB:

Celecoxib

Mn:

Number-average molecular weight

Mw:

Weight-average molecular weight

OA:

Osteoarthritis

PBS:

Phosphate buffered saline

PDI:

Polydispersity index

PGE2 :

Prostaglandin E2

PLGA-PTE:

Poly(lactic-co-glycolic acid) with poly(thioester) linkages

TA:

Triamcinolone acetonide

TNFα:

Tumor necrosis factor alpha

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Acknowledgments AND DISCLOSURES

This research forms part of the Project P2.01 IDiDAS of the research program of the BioMedical Materials institute, co-funded by the Dutch Ministry of Economic Affairs. The financial contribution of the Dutch Arthritis Association is gratefully acknowledged.

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Authors and Affiliations

  1. Department of Orthopaedics, University Medical Center Utrecht, Heidelberglaan 100, PO Box 85500, 3508 GA, Utrecht, The Netherlands

    Hsiao-yin Yang, Ruud Licht, Michiel Beekhuizen, Mattie van Rijen, F. Cumhur Öner, Wouter J. A. Dhert & Laura B. Creemers

  2. DSM Biomedical B.V., Geleen, The Netherlands

    Maarten van Dijk, Martina Källrot Janstål & Detlef Schumann

  3. Faculty of Veterinary Medicine, University of Utrecht, Utrecht, The Netherlands

    Wouter J. A. Dhert

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  1. Hsiao-yin Yang
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Correspondence to Laura B. Creemers.

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Yang, Hy., van Dijk, M., Licht, R. et al. Applicability of a Newly Developed Bioassay for Determining Bioactivity of Anti-Inflammatory Compounds in Release Studies − Celecoxib and Triamcinolone Acetonide Released from Novel PLGA-Based Microspheres . Pharm Res 32, 680–690 (2015). https://doi.org/10.1007/s11095-014-1495-z

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