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Objective This study aimed to develop a scale for nurses’ knowledge, attitudes and practices regarding the prevention and treatment of subcutaneous lipohypertrophy associated with insulin injections and evaluate its reliability and validity.Design A Delphi research and cross-sectional survey.Setting Medical, education and nursing experts from seven different regions conducted Delphi consultation by E-mail or Wechat, whereas clinical nurses from primary, secondary and tertiary general hospitals in different regions of Sichuan Province were conveniently selected for the questionnaire survey.Participants A total of 16 female experts aged 35–58 years participated in the Delphi expert consultation process; 812 clinical nurses participated in the questionnaire survey.Outcome measure Content validity, internal consistency reliability and construct validity of the scale.Results The scale comprises 3 dimensions and 36 items, with a content validity index of 0.99, a Cronbach’s α coefficient of 0.93, a split-half reliability of 0.87 and a test–retest reliability of 0.93. Exploratory factor analysis revealed a cumulative variance contribution rate of 53.38%, whereas confirmatory factor analysis indicated a χ2 degree of freedom 2 ratios of 2.65, a goodness-of-fit index of 0.86, a root mean square error of approximation of 0.06, a root mean square error of 0.04 and a comparative fit index of 0.90.Conclusion The developed scale demonstrated strong reliability and validity, enabling the assessment of nurses’ knowledge, attitudes and practices regarding the prevention and treatment of lipohypertrophy associated with insulin injections.