Review article
Improved in vitro models for preclinical drug and formulation screening focusing on 2D and 3D skin and cornea constructs

https://doi.org/10.1016/j.ejpb.2017.11.014Get rights and content

Abstract

The present overview deals with current approaches for the improvement of in vitro models for preclinical drug and formulation screening which were elaborated in a joint project at the Center of Pharmaceutical Engineering of the TU Braunschweig. Within this project a special focus was laid on the enhancement of skin and cornea models. For this reason, first, a computation-based approach for in silico modeling of dermal cell proliferation and differentiation was developed. The simulation should for example enhance the understanding of the performed 2D in vitro tests on the antiproliferative effect of hyperforin. A second approach aimed at establishing in vivo-like dynamic conditions in in vitro drug absorption studies in contrast to the commonly used static conditions. The reported Dynamic Micro Tissue Engineering System (DynaMiTES) combines the advantages of in vitro cell culture models and microfluidic systems for the emulation of dynamic drug absorption at different physiological barriers and, later, for the investigation of dynamic culture conditions. Finally, cryopreserved shipping was investigated for a human hemicornea construct. As the implementation of a tissue-engineering laboratory is time-consuming and cost-intensive, commercial availability of advanced 3D human tissue is preferred from a variety of companies. However, for shipping purposes cryopreservation is a challenge to maintain the same quality and performance of the tissue in the laboratory of both, the provider and the customer.

Introduction

This overview focuses on current approaches for the preclinical drug and formulation screening with improved in vitro models pursued within the joint research project SynFoBiA (Novel Synthesis and Formulation Methods for Poorly Soluble Drugs and Sensitive Biopharmaceuticals) at the Center of Pharmaceutical Engineering of the TU Braunschweig. Special focus was laid on in silico and in vitro skin models as well as on in vitro cornea models.

In the past preclinical testing of a novel compound – preferably as an aqueous solution – was usually carried out on excised tissue of an animal and in later stages on the animal itself, before the first clinical phase started in healthy human subjects. Nowadays, the use of cell culture and 2D as well as 3D tissue models is gaining increasing interest. However, the water solubility of novel substances is often extremely low [1] and requires sophisticated formulation approaches for dissolution and/or solubilization at an appropriate concentration at the right administration site [2], [3]. In addition, the number of animals used in experimentation is high causing many disadvantages such as high costs of laboratory animal care, ethical implications with regard to animal welfare in experiments, which cause the animal pain, suffering, injury, fear, or significantly disturb their general condition, and finally the issue of transferability of the results from animal to human, just to name a few. Against this background, the aim of replacing animal experiments whenever possible, of reducing the number of animal experiments, or at least of refining animal experiments as far as possible (3R principle of Russell and Burch, formulated as early as 1959 [4]) paves the way to alternative methods and their improved application.

Although a complete dispense of animal testing is actually agreed not to be possible, human cell cultivation provides tissue models which represent human three-dimensional (3D) organotypic constructs appropriate for replacing the respective animal tissue [5]. In many cases and with regard to specific issues, e.g., studying cell responses to drugs in terms of positive or negative effects on cell proliferation or toxicity, even two-dimensional (2D) human organotypic constructs are appropriate models. An excellent supplementation of cell-based models is provided by in silico models which focus on computation-based approaches [6]. Since the cultivation of human tissue is laborious, time-consuming and requires specific expertise in tissue engineering as well as sophisticated laboratory equipment, start-up companies have specialized in research and development, validation, manufacturing and commercialization of advanced human tissue. They provide ready-to-use tissue of a variety of organs cultivated under static conditions. For shipping purposes, this tissue has to be preserved in its ready-to-use state to perform the intended experiments with the customer’s laboratory. In this context, the cryopreservation of tissue for shipping purposes is a challenge to maintain the same quality and performance of the tissue in the lab of both the provider and the customer [7].

Following an introductory section with general considerations on 2D and 3D organotypic cell constructs/tissue models, the present overview starts with computation-based approaches to cell proliferation and differentiation of cultivated dermal tissue. The modeling was based on the in vitro 2D epithelial construct cultivation in the presence of hyperforin as a poorly soluble active pharmaceutical ingredient (API). The effects of hyperforin and hyperforin formulations on tissue models of the skin are presented in the second section, while the third section deals with improved drug absorption models for preclinical formulation testing. The improvement includes a better correspondence to the complex in vivo situation with regard to dynamic steady state conditions [8]. In this context, blood flow and/or dilution of donor and receiver compartments for example in pharmacokinetic studies of drug formulations are adjusted to the in vivo situation. Finally the cryopreservation of tissue for shipping purposes is the topic of the last section.

Section snippets

General considerations

Since the establishment of modern methods of cell cultivation and tissue engineering at the end of the 20th century, the number of research approaches for the development of in vitro alternatives has increased considerably. In addition to a large number of pharmacological and toxicological cellular assays, cell culture models have also been developed, which allow simple predictions of the pharmacokinetic behavior of active substances. Besides models for hepatic metabolism [9] or renal excretion

Conclusion

The presented overview covers successful examples of improved in vitro and in silico models for preclinical drug and formulation screening, starting with the usefulness of in silico modeling of “wound” closure processes and the positive or negative effects of poorly soluble drug molecules – as an example the focus was laid on hyperforin from the plant St. John’s Wort – on cell proliferation of tissue-engineered 2D and 3D skin. It further demonstrates the excellent performance of

Acknowledgment

The authors gratefully acknowledge funding by the Niedersächsisches Ministerium für Wissenschaft und Kultur within the joint research project SynFoBiA (Novel Synthesis and Formulation Methods for Poorly Soluble Drugs and Sensitive Biopharmaceuticals).

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