In recent years, hydrogels have emerged as premier platforms for drug delivery, attracting significant attention due to their exceptional biocompatibility and precisely tunable physical properties. Among the diverse crosslinking strategies available, enzymatic crosslinking mediated by microbial transglutaminase (mTG) offers distinctive advantages, particularly in enabling precise control over mechanical properties through gradual network formation. This study unveils the remarkable potential of mTG-mediated hydrogels for sophisticated drug delivery applications by harnessing their dynamic crosslinking characteristics. Our findings reveal that higher mTG concentrations generate more densely crosslinked networks with increased stiffness (up to 82.18 kPa at 10 mg/mL mTG), which surprisingly accelerates drug release kinetics by exerting pressure on encapsulated compounds. Conversely, lower mTG concentrations produce more permissive matrices with greater swelling capacity, facilitating sustained drug retention and controlled release profiles. Through systematic investigation of mTG concentration effects on mechanical properties, swelling behavior, and drug release patterns, we demonstrate unprecedented ability to fine-tune release kinetics with exceptional precision. Biocompatibility assessments confirm excellent cell viability across all formulations (80-98%), establishing these systems as highly promising for therapeutic applications. This research positions mTG-mediated hydrogels as next-generation drug delivery platforms capable of achieving customized release profiles through simple enzymatic modulation, offering transformative potential for targeted therapeutics. Future investigations will expand this versatile system's applicability to diverse bioactive compounds and evaluate its performance under complex physiological conditions.

Abstract
1. Introduction
2. Materials and Methods
    2.1. Materials
    2.2. Hydrogel preparation
    2.3. Mechanical properties testing
    2.4. Drug encapsulation and release study
    2.5. Swelling ratio measurement
    2.6. Cell viability assay
    2.7. Statistical analysis
3. Results and Discussion
    3.1. Preparation of enzymaticallycrosslinked hydrogel
    3.2. Hydrogel stiffness
    3.3. Drug release profiles
    3.4. Swelling ratios
    3.5. Cell viability
    3.6. Discussion
4. Conclusions
Acknowlegements
References

• Kyubae Lee(Department of Biomedical Materials, Konyang University, 158, Gwanjeodong-ro, Seo-gu, Daejeon, 35365, Republic of Korea) Corresponding author